LOCUS       pET28a-pT5-IF2          7933 bp    DNA     circular SYN 22-JUL-2025
DEFINITION  Nucleus PURE: pET28a-pT5-IF2 in pET28a protein expression plasmid.
            See https://nucleus.bnext.bio/ for more details..
ACCESSION   pET28a-pT5-IF2
VERSION     pET28a-pT5-IF2
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     rep_origin      77..665
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     promoter        77..105
                     /label="RNAII promoter"
                     /note="Color: #ffffff NCBI Feature Key: promoter NCBI Join
                     Type: join Transferred From: RNAII_promoter Transferred
                     Similarity: 100.00%"
     misc_RNA        113..665
                     /label="RNAII transcript (complementary strand)"
                     /note="RNAII transcript (complementary strand)"
     misc_RNA        complement(116..223)
                     /label="RNAI transcript"
                     /note="RNAI transcript"
     promoter        complement(230..259)
                     /label="RNA I promoter"
                     /note="Color: #ffffff NCBI Feature Key: promoter NCBI Join
                     Type: join Transferred From: RNA_I_promoter Transferred
                     Similarity: 100.00%"
     terminator      669..712
                     /label="rop/bom terminator"
     misc_binding    752..760
                     /label="dnaA binding site"
                     /bound_moiety="dnaA"
     misc_feature    785..848
                     /label="L-strand Y effector site"
                     /note="L-strand Y effector site"
     misc_feature    851..990
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     misc_feature    1031..1187
                     /label="H-strand Y effector site"
                     /note="H-strand Y effector site"
     CDS             complement(1092..1283)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein"
                     /label="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     RBS             1284..1293
     misc_feature    complement(1359..1377)
                     /label="ROP calculated promoter"
     CDS             complement(2092..3174)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label="lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(3175..3252)
                     /label="lacI promoter"
                     /note="Color: #ffffff NCBI Feature Key: promoter
                     Transferred From: lacI_promoter Transferred Similarity:
                     100.00%"
     source          3561..6378
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        3561..3605
                     /label="T5 promoter"
     protein_bind    3581..3597
                     /label="lac operator"
     protein_bind    3613..3629
                     /label="lac operator"
     RBS             3648..3659
                     /label="RBS"
     CDS             3666..3677
                     /codon_start=1
                     /label="N-term"
                     /translation="MRGS"
     CDS             3678..3695
                     /codon_start=1
                     /product="HHHHHH"
                     /label="6xHis"
                     /translation="HHHHHH"
     CDS             3696..3701
                     /codon_start=1
                     /label="Linker"
                     /translation="GS"
     CDS             3702..6371
                     /codon_start=1
                     /label="IF2"
                     /translation="MTDVTIKTLAAERQTSVERLVQQFADAGIRKSADDSVSAQEKQTL
                     IDHLNQKNSGPDKLTLQRKTRSTLNIPGTGGKSKSVQIEVRKKRTFVKRDPQEAERLAA
                     EEQAQREAEEQARREAEESAKREAQQKAEREAAEQAKREAAEQAKREAAEKDKVSNQQD
                     DMTKNAQAEKARREQEAAELKRKAEEEARRKLEEEARRVAEEARRMAEENKWTDNAEPT
                     EDSSDYHVTTSQHARQAEDESDREVEGGRGRGRNAKAARPKKGNKHAESKADREEARAA
                     VRGGKGGKRKGSSLQQGFQKPAQAVNRDVVIGETITVGELANKMAVKGSQVIKAMMKLG
                     AMATINQVIDQETAQLVAEEMGHKVILRRENELEEAVMSDRDTGAAAEPRAPVVTIMGH
                     VDHGKTSLLDYIRSTKVASGEAGGITQHIGAYHVETENGMITFLDTPGHAAFTSMRARG
                     AQATDIVVLVVAADDGVMPQTIEAIQHAKAAQVPVVVAVNKIDKPEADPDRVKNELSQY
                     GILPEEWGGESQFVHVSAKAGTGIDELLDAILLQAEVLELKAVRKGMASGAVIESFLDK
                     GRGPVATVLVREGTLHKGDIVLCGFEYGRVRAMRNELGQEVLEAGPSIPVEILGLSGVP
                     AAGDEVTVVRDEKKAREVALYRQGKFREVKLARQQKSKLENMFANMTEGEVHEVNIVLK
                     ADVQGSVEAISDSLLKLSTDEVKVKIIGSGVGGITETDATLAAASNAILVGFNVRADAS
                     ARKVIEAESLDLRYYSVIYNLIDEVKAAMSGMLSPELKQQIIGLAEVRDVFKSPKFGAI
                     AGCMVTEGVVKRHNPIRVLRDNVVIYEGELESLRRFKDDVNEVRNGMECGIGVKNYNDV
                     RTGDVIEVFEIIEIQRTIA"
     terminator      6442..6489
                     /label="T7 terminator"
                     /note="Color: #ffffff NCBI Feature Key: terminator
                     Transferred From: T7_terminator Transferred Similarity:
                     100.00%"
     rep_origin      6526..6981
                     /direction=RIGHT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(7073..7888)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label="KanR"
                     /note="confers resistance to kanamycin"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
ORIGIN
        1 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
       61 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
      121 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      181 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
      241 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      301 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      361 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      421 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      481 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      541 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      601 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      661 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      721 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
      781 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
      841 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
      901 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
      961 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     1021 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     1081 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
     1141 agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc
     1201 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
     1261 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg
     1321 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat
     1381 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag
     1441 agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag
     1501 ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc
     1561 cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc
     1621 gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc
     1681 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg
     1741 cgcacccgtg gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg
     1801 gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac
     1861 aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct
     1921 gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata
     1981 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     2041 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     2101 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     2161 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     2221 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     2281 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     2341 gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc
     2401 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     2461 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     2521 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     2581 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     2641 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     2701 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     2761 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     2821 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     2881 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     2941 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     3001 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     3061 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     3121 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     3181 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
     3241 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc
     3301 ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga
     3361 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     3421 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     3481 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg
     3541 agatctcgat cccgcgaaat tcataaaaaa tttatttgct ttgtgagcgg ataacaatta
     3601 taatagattc aattgtgagc ggataacaat ttcacacaga attcattaaa gaggagaaat
     3661 taactatgag aggatcgcat caccatcacc atcacggatc catgacagat gtaacgatta
     3721 aaacgctggc cgcagagcga cagacctccg tggaacgcct ggtacagcaa tttgctgatg
     3781 caggtatccg gaagtctgct gacgactctg tgtctgcaca agagaaacag actttgattg
     3841 accacctgaa tcagaaaaat tcaggcccgg acaaattgac gctgcaacgt aaaacacgca
     3901 gcacccttaa cattcctggt accggtggaa aaagcaaatc ggtacaaatc gaagtccgca
     3961 agaaacgcac ctttgtgaaa cgcgatccgc aagaggctga acgccttgca gcggaagagc
     4021 aagcgcagcg tgaagcggaa gagcaagccc gtcgtgaggc agaagaatcg gctaaacgcg
     4081 aggcgcaaca aaaagctgaa cgtgaggccg cagaacaagc taagcgtgaa gctgctgaac
     4141 aagcgaaacg tgaagctgcg gaaaaagaca aagtgagcaa tcaacaagac gatatgacta
     4201 aaaacgccca ggctgaaaaa gcccgccgtg agcaggaagc tgcagagctc aagcgtaaag
     4261 ctgaagaaga agcgcgtcgt aaactcgaag aagaagcacg tcgcgttgct gaagaagcac
     4321 gtcgtatggc ggaagaaaac aaatggactg ataacgcgga accgactgaa gattccagcg
     4381 attatcacgt cactacttct caacatgctc gccaggcaga agacgaaagc gatcgtgaag
     4441 tcgaaggcgg ccgtggccgt ggtcgtaacg cgaaagcagc gcgtccgaag aaaggcaaca
     4501 aacacgctga atcaaaagct gatcgtgaag aagcacgcgc agcagtacgt ggcggtaaag
     4561 gcggaaaacg taaaggttct tcgctgcagc aaggcttcca gaagcctgct caggccgtta
     4621 accgtgacgt tgtgatcggc gaaactatca ccgttggcga actggcgaac aagatggcgg
     4681 ttaaaggctc tcaggtcatc aaagcgatga tgaaactggg cgcaatggca accatcaacc
     4741 aggttatcga tcaggaaacc gcacagctgg ttgctgaaga gatgggccat aaagttatcc
     4801 tgcgtcgtga aaacgagctg gaagaggcgg taatgagcga ccgtgacacg ggtgctgcgg
     4861 ctgaaccgcg cgcgccggtt gtgaccatca tgggtcacgt tgaccacggt aaaacctctc
     4921 tgctggacta cattcgttca acgaaagtgg cctctggcga agcgggcggc attacccagc
     4981 acattggtgc ataccacgtt gaaactgaaa acggcatgat caccttcctg gacaccccgg
     5041 ggcacgccgc gtttacttca atgcgtgctc gtggtgcgca ggcaacggac atcgtagtcc
     5101 tggttgttgc tgccgacgac ggtgtgatgc cgcagaccat cgaagcaatc cagcacgcga
     5161 aagcggcgca ggtaccggtg gtggttgcag tgaacaagat cgataaacca gaagctgatc
     5221 cggatcgcgt taagaacgaa ctctcccagt acggcatcct gccggaagag tggggcggtg
     5281 aaagccagtt cgtacacgta tctgcgaaag cgggtaccgg tatcgatgaa ctgctggacg
     5341 ctatcctgct gcaggcggaa gttctggagc tgaaagcggt acgtaaaggt atggcgagcg
     5401 gtgcggttat cgaatccttc ctcgataaag gtcgtggtcc ggttgctacc gttctggtac
     5461 gtgaaggtac tctgcacaag ggcgatatcg ttctgtgtgg cttcgaatac ggtcgtgttc
     5521 gtgcgatgcg taacgaactg ggtcaggaag tgctggaagc gggtccgtcc attccggtgg
     5581 aaatcctcgg cctgtccggc gtaccggctg cgggtgatga agttaccgtt gtacgtgacg
     5641 agaagaaagc gcgtgaagtt gcactctatc gtcagggtaa attccgcgaa gttaaactgg
     5701 cgcgtcagca gaaatctaaa ctcgagaaca tgttcgccaa catgaccgaa ggcgaagttc
     5761 acgaagtgaa tatcgtcctg aaggcagacg tacagggttc tgtcgaagcg atctccgact
     5821 ccttgctgaa actgtctact gacgaagtta aagtgaagat catcggttct ggcgtaggtg
     5881 gtatcaccga aaccgacgcc accctggctg cggcgtccaa cgccatcctg gttggcttta
     5941 acgtacgtgc tgatgcctct gcacgtaaag tgattgaagc ggaaagcctg gatctgcgtt
     6001 actactccgt catctataac ctgattgacg aagtgaaagc ggcgatgagc ggtatgctgt
     6061 ctccggaact gaaacagcag attatcggtc tggcggaagt tcgtgacgtg ttcaaatcgc
     6121 cgaaatttgg tgccatcgca ggctgtatgg ttaccgaagg tgtggttaaa cgtcacaacc
     6181 cgatccgcgt tctgcgtgac aacgtggtta tctacgaagg cgagctggag tccctgcgcc
     6241 gcttcaaaga tgacgttaac gaagtccgta acggtatgga atgtggtatc ggcgttaaga
     6301 actacaacga cgtccgcact ggcgatgtga tcgaagtatt cgaaatcatc gagatccaac
     6361 gtaccattgc ttaagcttga tccggctgct aacaaagccc gaaaggaagc tgagttggct
     6421 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     6481 ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg
     6541 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     6601 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     6661 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     6721 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     6781 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     6841 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     6901 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     6961 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     7021 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaat tcttagaaaa
     7081 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt
     7141 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     7201 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     7261 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
     7321 gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac
     7381 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
     7441 cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
     7501 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
     7561 atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag
     7621 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
     7681 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
     7741 gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc
     7801 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc
     7861 aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag
     7921 acagttttat tgt
//
