LOCUS       sequence-13267-e        8136 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8136)
  AUTHORS   Gumbiner-Russo LM, Lombardo MJ, Ponder RG, Rosenberg SM
  TITLE     The TGV transgenic vectors for single-copy gene expression from the 
            Escherichia coli chromosome.
  JOURNAL   Gene. 2001 Jul 25. 273(1):97-104.
  PUBMED    11483365
REFERENCE   2  (bases 1 to 8136)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene. 2001 
            Jul 25. 273(1):97-104."
COMMENT     Sequence Label: pTGV-Tet SMR5970
FEATURES             Location/Qualifiers
     source          1..8136
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(2187..3377)
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     3006..3025
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        complement(join(3425..3430,3431..3447,3448..3453))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     protein_bind    3700..3721
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3736..3741,3742..3759,3760..3766)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3774..3790
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by 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)."
     primer_bind     3779..3801
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3798..3814
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3798..3814
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3832..3850
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3832..3849
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     rep_origin      complement(6328..6916)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6408..6427)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7087..7878,7879..7947))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7710..7729
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7948..8052)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(8120..8136,1..2)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gccctggcgc gcctaggcca tatgggcccg ggcgagcata ttgatccgct gcaaactgaa
       61 attggcagca tggtctatgc ggtgaaacca ggcgatggtt ctgcgcgtga acaggcggcg
      121 agctgccagc gtgtgattgg cggtctggcg aatattgccg aggagtacgc gactaaacgc
      181 tatcgttcta acgtcatcaa ctgggggatg ttaccgctgc aaatggcgga agtaccaacc
      241 tttgaagtgg gggattacat ttacatccct ggcattaaag cggcgctgga taatccgggt
      301 acgacgttta aaggttatgt gatccatgaa gatgcgccgg taacggaaat tacgctctat
      361 atggaaagtc tgactgctga agagcgcgag attatcaagg cgggtagttt gattaacttc
      421 aataaaaacc gtcagatgta aaaagcgcca tgtgaatgta ggtcgcattc ggcacttatt
      481 gtcggatgcg atgcttgcgc atcttatccg acctacgaat cgcatcgaat ctgtaggcca
      541 gataaggcat tttcgcagca tccggcactt attgtcggat gcgatgcttg cgcatcttat
      601 ccgacctaca aatcgcatcg aaccgtaggc cggataaggc gtttacgccg catccggcaa
      661 atagttaatt gctcttactt cttcgcctct gcaaccactt tactacccac gccgcggtta
      721 ttgtattccc acatgcggtt gtagttagtg tcattcagat tgcgctgtat ttcgtcgtta
      781 tcatctacgc tgccggtatt acccgcaaac ggacgattag agatcaccgc atcggcccac
      841 ggtttagccg tgttaaaacc ttcgttgatg gcgctatcac ggatcaccac ctgaccgttg
      901 gtattggcat caacatccag cgagcggccc agttgcgcca caccatcacc gaaagcattg
      961 aaacggctgt ttacggcgag gaaaccgtag taaatgttgg acagcgtagc cggtgcaaac
     1021 acatacgctt cttgctgagt acgtgagttc accacgcgga attcggtgtt atcgaacacc
     1081 actgcgccgc gaccagaaac gatatccaca tccccttcaa tgtagctgtt ggtcaccagc
     1141 gtacgcggct gacgattcgt ttccagacgg ttctgcacac cgctgttggt gacaaagaag
     1201 gtgttctgac gaccgagaat gttaacgttg ttaatctgta cctggtcacc atcagtacgc
     1261 agtgccaccg ccggatggtt acctgcatct acgctatcgc ccagcgtgtt ttcgatggtc
     1321 agattttgca gttgcaggcc attgttttgt gaccagaaga ccgcagagca gagaacaccg
     1381 atactgtcgc tgcgtttgct ctggcagcta tcgtacatat accacgctgg tttacctggc
     1441 atatatttgc cgcgcgggtt gacgtcgtga cgccagtcgg cagggctcat gccaccatca
     1501 agggaaagcc caatcttcac atcaatcggt ttttcacctg taccgtacag agtaattcca
     1561 cccggagcgg cagggacata taccgttccc tgatactcac caggcatcac ggcaatatac
     1621 tggcgcttgt tggtacgctt gataattgcc gcatctaccg ccgcctgaat cgtggtatgc
     1681 gttacacctt gagtgcccgc cgggccgaca acaaagtcag gttgcgcagg cagggtaatc
     1741 ggggaaggat tccacgctgc agcacctggt gtcagggatg caaaatagtg ttgagcatcg
     1801 aaattctgcg cttcttttgc cgacagaatc gggcgagaag aggtaccagg cgcggtttga
     1861 tcagaaggac gttgatcggg cggggttgag ctacaggcgg tcagcgtcac gccaaaagcc
     1921 aatgccagcg ccagacggga aactgaaaat gtgttcacag gttgctccgg gctatgaaat
     1981 agaaaaatga atccgttgaa ggcatgcaag cttgatccag gacccaacgc tgcccgagat
     2041 gcgccgcgtg cggctgctgg agatggcgga cgcgatggat atgttctgcc aagggttggt
     2101 ttgcgcattc acagttctcc gcaagaattg attggctcca attcttggag tggtgaatcc
     2161 gttagcgagg tgccgccggc ttccattcag gtcgaggtgg cccggctcca tgcaccgcga
     2221 cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta caatccatgc caacccgttc
     2281 catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca gcggtccagt gatcgaagtt
     2341 aggctggtaa gagccgcgag cgatccttga agctgtccct gatggtcgtc atctacctgc
     2401 ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag aagaatcata
     2461 atggggaagg ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg
     2521 gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg
     2581 acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc
     2641 gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt
     2701 cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc
     2761 gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg gcgctctccc
     2821 ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc
     2881 gccgcaagga atggtgcatg taaggagatg gcgcccaaca gtcccccggc cacggggcct
     2941 gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc
     3001 ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg
     3061 gccacgatgc gtccggcgta gagaatccac aggacgggtg tggtcgccat gatcgcgtag
     3121 tcgatagtgg ctccaagtag cgaagcgagc aggactgggc ggcggccaaa gcggtcggac
     3181 agtgctccga gaacgggtgc gcatagaaat tgcatcaacg catatagcgc tagcagcacg
     3241 ccatagtgac tggcgatgct gtcggaatgg acgatatccc gcaagaggcc cggcagtacc
     3301 ggcataacca agcctatgcc tacagcatcc agggtgacgg tgccgaggat gacgatgagc
     3361 gcattgttag atttcataca cggtgcctga ctgcgttagc aatttaactg tgataaacta
     3421 ccgcattaaa gctaatcgat gataagctgt caaacatgag aattaattca tgttctttcc
     3481 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     3541 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     3601 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     3661 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     3721 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     3781 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc tatttaggtg
     3841 acactataga atactcaagc ttgcatgcgt ctagagctcg aggtagatct gaaagcttga
     3901 atcgatgtac gtactatacc ggtgcgaaac gggaaggtaa aaagacaaaa agttgttttt
     3961 aataccttta agtgatacca gatggcattg cgccatctgg cagagtgatt aactaaacat
     4021 cgcagtaatc gaggcgcttg ccagagagtg gaaatgaacg ttaaacccga ccatcgcgcc
     4081 gctggcacct tcatcgacat caatacgttc tatatccagc gcgtgaacgg taaaaatgta
     4141 gcgatgagtt tcgcctttcg gcggtgctgc gccatcgtac ccggttttac caaagtcggt
     4201 acgcgtctgc aaaacgccgt ctggcattgc taccagacca gagccaaacc cttgcggtaa
     4261 tacgcgggta tcagcgggta agttaacaac tacccagtgc caccagccgg agccggttgg
     4321 cgcatccggg tcgtagcagg tgacaacaaa acttttcgtt cccgcaggaa catcatccca
     4381 cgccagatgc ggtgaaatat tatcgccatc gtaacccatg ccgttaaaga catgacgatg
     4441 cggcaattta tcgccatcgc gcagatcgtt actgatgagt ttcatgaacc ctcctttctt
     4501 gtttgcagaa agtgtagcca gaaaccctca cgcggacttc tcgttattgg caaaaaaatg
     4561 tttcatcctg taccgcgcgg ttaaccgctg cggtcagacg ctgcaactgt tgcgggagaa
     4621 taatataggg cggcatcagg taaatcagtt tgccaaaagg ccggatccag acaccctgtt
     4681 cgacaaagaa tttttgcagc gccgccatat tcaccggatg agtggtttcg accacgccaa
     4741 tggcccccag tacgcgcaca tcggcaacca tttcggcatc acgggcgggg gcaagttgct
     4801 cgcgcagctg tacttcaata tccgccacct gttgctgcca gtcgccagat tcgagaatcg
     4861 ccaggctggc gtttgctgcc gcgcaggcca gcggattgcc cataaaagtt ggcccatgca
     4921 taaagcaacc ggcttcaccg ttactgatgg tttctgcaac ctcgcgcgtg gtgagtgtgg
     4981 cggaaagggt cattgtgccg ccggttaagg ctttaccgag gcacaaaatg tccggcgcga
     5041 tttctgcatg ttcacaggca aacagtttcc cggtacgacc aaatccagtg gcgatctcgt
     5101 cggcaatcag caagatacct tcgcgatcgc atattttgcg gattcgtttt aaccattccg
     5161 gatggtacat gcgcatcccg cctgcgccct ggacaatcgg ctcaatgatc accgccgcga
     5221 tttcatgacg atgcgccgcc atcaggcggg caaagcccac catatcgcgc tcatcccatt
     5281 cgccatccat gcggctttgc ggggcgggag caaacaggtt ttctggcagg tagcctttcc
     5341 acagactgtg cattgagtta tccggatcgc acaccgacat cgcgccaaag gtatcgccat
     5401 gataaccatt gcggaaggtc agaaaacgct ggcgcgcttc gcctttggct tgccagtact
     5461 gcaacgccat tttcatcgcc acttccaccg ctacggaacc ggagtccgcg agaaaaacgc
     5521 actccagcgg ttgcggcgtc atcgccacca gtttgcggca cagctcaatg gctggcgcat
     5581 gggtgatacc gccaaacatc acatgcgaca tggcatcaat ttgcgacttc atcgccgcat
     5641 taagctgcgg gtgattgtag ccgtggatcg ccgcccacca ggacgacata ccgtcaacca
     5701 ggcgtctgcc gtcagacaaa atcagctcgc aaccttcggc gctcaccacc ggataaaccg
     5761 gcagagggga ggtcatggat gtgtatgggt gccagatatg gcgttggtca aaggcaagat
     5821 cgtccgttgt cataatcgac ttgtaaacca aattgaaaag atttaggttt acaagtctac
     5881 accgaattaa caacaaaaaa cacgttttgg agaagcccca tggctcaccg cccacgctgg
     5941 acattgtcgc aagtcacaga attatttgaa aaaccgttgc tggatctgct gtttgaagcg
     6001 cagcaggtgc atcgccagca tttcgatcct cgtcaggtgc aggtcagcac gttgctgcat
     6061 atggcgcgcc taggccaatt gggcccgggc atatgggtgt gaaataccgc acagatgcgt
     6121 aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6181 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6241 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6301 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6361 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6421 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6481 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6541 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6601 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6661 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6721 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6781 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6841 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6901 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6961 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7021 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7081 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7141 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7201 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7261 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7321 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7381 gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7441 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7501 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7561 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7621 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7681 gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca gaactttaaa
     7741 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7801 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7861 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7921 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7981 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     8041 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     8101 catgacatta acctataaaa ataggcgtat cacgag
//
