LOCUS       Exported                3719 bp DNA     circular SYN 10-SEP-2025
DEFINITION  Bacterial vector with a ColA origin for the co-expression of two 
            genes.
ACCESSION   .
VERSION     .
KEYWORDS    pCOLADuet-1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3719)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 3719)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 3719)
  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..3719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          join(44..3719,1..43)
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..45
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    46..70
                     /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             85..107
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             114..116
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             126..143
                     /label=6xHis
                     /note="6xHis affinity tag"
     promoter        257..275
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    276..300
                     /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             329..334
                     /note="ribosome binding site"
     CDS             343..345
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             409..453
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     terminator      505..552
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(782..1597)
                     /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(1598..1689)
                     /label=AmpR promoter
     rep_origin      complement(1707..2342)
                     /direction=LEFT
                     /label=ColA ori
                     /note="Plasmids containing the ColA origin of replication
                     can be propagated in E. coli cells that contain additional 
                     plasmids with compatible origins."
     protein_bind    complement(2506..2527)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(2540..3622)
                     /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(3623..3700)
                     /label=lacI promoter
ORIGIN
        1 cttatgcgac tcctgcatta ggaaattaat acgactcact ataggggaat tgtgagcgga
       61 taacaattcc cctgtagaaa taattttgtt taactttaat aaggagatat accatgggca
      121 gcagccatca ccatcatcac cacagccagg atccgaattc gagctcggcg cgcctgcagg
      181 tcgacaagct tgcggccgca taatgcttaa gtcgaacaga aagtaatcgt attgtacacg
      241 gccgcataat cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc
      301 ccatcttagt atattagtta agtataagaa ggagatatac atatggcaga tctcaattgg
      361 atatcggccg gccacgcgat cgctgacgtc ggtaccctcg agtctggtaa agaaaccgct
      421 gctgcgaaat ttgaacgcca gcacatggac tcgtctacta gcgcagctta attaacctag
      481 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
      541 aggggttttt tgctgaaacc tcaggcattt gagaagcaca cggtcacact gcttccggta
      601 gtcaataaac cggtaaacca gcaatagaca taagcggcta tttaacgacc ctgccctgaa
      661 ccgacgacaa gctgacgacc gggtctccgc aagtggcact tttcggggaa atgtgcgcgg
      721 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgaattaatt
      781 cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa
      841 taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc
      901 ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac
      961 ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga
     1021 ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc
     1081 agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt
     1141 gcgcctgagc gagacgaaat acgcggtcgc tgttaaaagg acaattacaa acaggaatcg
     1201 aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat
     1261 attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat
     1321 catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt
     1381 ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa
     1441 acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga
     1501 cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg
     1561 gcctagagca agacgtttcc cgttgaatat ggctcatact cttccttttt caatattatt
     1621 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     1681 ataaacaaat aggcatgcta gcgcagaaac gtcctagaag atgccaggag gatacttagc
     1741 agagagacaa taaggccgga gcgaagccgt ttttccatag gctccgcccc cctgacgaac
     1801 atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc
     1861 aggcgtttcc ccctgatggc tccctcttgc gctctcctgt tcccgtcctg cggcgtccgt
     1921 gttgtggtgg aggctttacc caaatcacca cgtcccgttc cgtgtagaca gttcgctcca
     1981 agctgggctg tgtgcaagaa ccccccgttc agcccgactg ctgcgcctta tccggtaact
     2041 atcatcttga gtccaacccg gaaagacacg acaaaacgcc actggcagca gccattggta
     2101 actgagaatt agtggattta gatatcgaga gtcttgaagt ggtggcctaa cagaggctac
     2161 actgaaagga cagtatttgg tatctgcgct ccactaaagc cagttaccag gttaagcagt
     2221 tccccaactg acttaacctt cgatcaaacc gcctccccag gcggtttttt cgtttacaga
     2281 gcaggagatt acgacgatcg taaaaggatc tcaagaagat cctttacgga ttcccgacac
     2341 catcactcta gatttcagtg caatttatct cttcaaatgt agcacctgaa gtcagcccca
     2401 tacgatataa gttgtaattc tcatgttagt catgccccgc gcccaccgga aggagctgac
     2461 tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact
     2521 tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     2581 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg
     2641 tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag
     2701 agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg
     2761 tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga
     2821 tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct
     2881 gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt
     2941 gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat
     3001 tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg
     3061 ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc
     3121 gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa
     3181 gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca
     3241 gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt
     3301 tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat
     3361 cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg
     3421 tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa
     3481 tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc
     3541 tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat
     3601 cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc
     3661 atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctcc
//
