LOCUS       Exported                3829 bp DNA     circular SYN 10-SEP-2025
DEFINITION  Bacterial vector with an RSF 1030 origin for the co-expression of 
            two genes.
ACCESSION   .
VERSION     .
KEYWORDS    pRSFDuet-1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3829)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 3829)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 3829)
  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..3829
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          join(48..3829,1..47)
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        31..49
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    50..74
                     /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             89..111
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             118..120
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             130..147
                     /label=6xHis
                     /note="6xHis affinity tag"
     promoter        261..279
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    280..304
                     /label=lac repressor encoded by lacI binding site
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="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             333..338
                     /note="ribosome binding site"
     CDS             347..349
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             413..457
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     terminator      509..556
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(786..1601)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        complement(1602..1693)
                     /label=AmpR promoter
     rep_origin      complement(1709..2458)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     protein_bind    complement(2620..2641)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(2654..3736)
                     /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(3737..3814)
                     /label=lacI promoter
ORIGIN
        1 ctcccttatg cgactcctgc attaggaaat taatacgact cactataggg gaattgtgag
       61 cggataacaa ttcccctgta gaaataattt tgtttaactt taataaggag atataccatg
      121 ggcagcagcc atcaccatca tcaccacagc caggatccga attcgagctc ggcgcgcctg
      181 caggtcgaca agcttgcggc cgcataatgc ttaagtcgaa cagaaagtaa tcgtattgta
      241 cacggccgca taatcgaaat taatacgact cactataggg gaattgtgag cggataacaa
      301 ttccccatct tagtatatta gttaagtata agaaggagat atacatatgg cagatctcaa
      361 ttggatatcg gccggccacg cgatcgctga cgtcggtacc ctcgagtctg gtaaagaaac
      421 cgctgctgcg aaatttgaac gccagcacat ggactcgtct actagcgcag cttaattaac
      481 ctaggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
      541 cttgaggggt tttttgctga aacctcaggc atttgagaag cacacggtca cactgcttcc
      601 ggtagtcaat aaaccggtaa accagcaata gacataagcg gctatttaac gaccctgccc
      661 tgaaccgacg acaagctgac gaccgggtct ccgcaagtgg cacttttcgg ggaaatgtgc
      721 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgaatt
      781 aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
      841 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
      901 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
      961 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     1021 acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca
     1081 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
     1141 gattgcgcct gagcgagacg aaatacgcgg tcgctgttaa aaggacaatt acaaacagga
     1201 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     1261 ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat
     1321 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
     1381 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     1441 agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc
     1501 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     1561 cgcggcctag agcaagacgt ttcccgttga atatggctca tactcttcct ttttcaatat
     1621 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     1681 aaaaataaac aaataggcat gcagcgctct tccgcttcct cgctcactga ctcgctacgc
     1741 tcggtcgttc gactgcggcg agcggtgtca gctcactcaa aagcggtaat acggttatcc
     1801 acagaatcag gggataaagc cggaaagaac atgtgagcaa aaagcaaagc accggaagaa
     1861 gccaacgccg caggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1921 acgctcaagc cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1981 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2041 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgttggt atctcagttc
     2101 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2161 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2221 actggcagca gccattggta actgatttag aggactttgt cttgaagtta tgcacctgtt
     2281 aaggctaaac tgaaagaaca gattttggtg agtgcggtcc tccaacccac ttaccttggt
     2341 tcaaagagtt ggtagctcag cgaaccttga gaaaaccacc gttggtagcg gtggtttttc
     2401 tttatttatg agatgatgaa tcaatcggtc tatcaagtca acgaacagct attccgttac
     2461 tctagatttc agtgcaattt atctcttcaa atgtagcacc tgaagtcagc cccatacgat
     2521 ataagttgta attctcatgt tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt
     2581 gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt
     2641 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     2701 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc
     2761 ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt
     2821 gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta
     2881 acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg
     2941 caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt
     3001 tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa
     3061 aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag
     3121 tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg
     3181 ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac
     3241 cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata
     3301 acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat
     3361 agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg
     3421 cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc
     3481 gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa
     3541 cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat
     3601 tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct
     3661 ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata
     3721 acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca
     3781 taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgct
//
