LOCUS       Exported                5422 bp DNA     circular SYN 10-SEP-2025
DEFINITION  Bacterial vector for expression of N-terminally S-tagged proteins 
            with an enterokinase site. For other reading frames, use pET-30b(+) 
            or pET-30c(+).
ACCESSION   .
VERSION     .
KEYWORDS    pET-30a(+)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5422)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5422)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5422)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 10, 2025 from SnapGene 6.0.2
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          join(679..5422,1..678)
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        244..262
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    263..287
                     /label=lac operator
                     /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)."
     RBS             302..324
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             332..334
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             335..352
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             362..379
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             386..430
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     CDS             446..460
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     misc_feature    462..521
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             522..539
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      606..653
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      690..1145
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(1241..2053)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      2175..2763
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2949..3091
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3196..3384)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    4159..4180
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4196..5275)
                     /label=lacI
                     /note="lac repressor"
     promoter        5276..5353
                     /label=lacI promoter
ORIGIN
        1 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
       61 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
      121 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
      181 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatcgatctc gatcccgcga
      241 aattaatacg actcactata ggggaattgt gagcggataa caattcccct ctagaaataa
      301 ttttgtttaa ctttaagaag gagatataca tatgcaccat catcatcatc attcttctgg
      361 tctggtgcca cgcggttctg gtatgaaaga aaccgctgct gctaaattcg aacgccagca
      421 catggacagc ccagatctgg gtaccgacga cgacgacaag gccatggctg atatcggatc
      481 cgaattcgag ctccgtcgac aagcttgcgg ccgcactcga gcaccaccac caccaccact
      541 gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
      601 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
      661 gaggaactat atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
      721 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
      781 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
      841 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
      901 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
      961 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     1021 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     1081 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     1141 aatttcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     1201 aatacattca aatatgtatc cgctcatgaa ttaattctta gaaaaactca tcgagcatca
     1261 aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt
     1321 tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc
     1381 ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa
     1441 taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa
     1501 gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat
     1561 cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc
     1621 gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg
     1681 ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg
     1741 ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct
     1801 tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa
     1861 catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc
     1921 catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc
     1981 catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac gtttcccgtt
     2041 gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc
     2101 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     2161 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2221 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2281 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     2341 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2401 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2461 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     2521 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     2581 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     2641 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     2701 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     2761 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     2821 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     2881 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     2941 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     3001 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc
     3061 gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc
     3121 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     3181 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta
     3241 aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag
     3301 ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag
     3361 ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg
     3421 ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca
     3481 tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca
     3541 gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca
     3601 gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt
     3661 ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt
     3721 cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg
     3781 ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat
     3841 gcgcacccgt ggggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt
     3901 ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga
     3961 caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc
     4021 tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat
     4081 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
     4141 tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc
     4201 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4261 gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca
     4321 acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg
     4381 tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc
     4441 tgtcttcggt atcgtcgtat cccactaccg agatgtccgc accaacgcgc agcccggact
     4501 cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg
     4561 gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt
     4621 cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag
     4681 ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt
     4741 gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa
     4801 tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg
     4861 cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga
     4921 cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta
     4981 ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa
     5041 tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt
     5101 tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt
     5161 ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg
     5221 tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca
     5281 ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc
     5341 attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag
     5401 cccagtagta ggttgaggcc gt
//
