LOCUS       sequence-13265-e        8493 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 8493)
  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 8493)
  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-Express-Tet SMR5918
FEATURES             Location/Qualifiers
     source          1..8493
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(2208..3398)
                     /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     3027..3046
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        complement(join(3446..3451,3452..3468,3469..3474))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     protein_bind    3721..3742
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3757..3762,3763..3780,3781..3787)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3795..3811
                     /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     3800..3822
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3819..3835
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3819..3835
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3853..3871
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3853..3870
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        3914..4198
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     4121..4140
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     rep_origin      complement(6685..7273)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6765..6784)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7444..8235,8236..8304))
                     /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     8067..8086
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8305..8409)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(8477..8493,1..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 ggcatgtctg agctcctgtc atgcatgcaa gcttgatcca
     2041 ggacccaacg ctgcccgaga tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga
     2101 tatgttctgc caagggttgg tttgcgcatt cacagttctc cgcaagaatt gattggctcc
     2161 aattcttgga gtggtgaatc cgttagcgag gtgccgccgg cttccattca ggtcgaggtg
     2221 gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga caaggtatag ggcggcgcct
     2281 acaatccatg ccaacccgtt ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc
     2341 agcggtccag tgatcgaagt taggctggta agagccgcga gcgatccttg aagctgtccc
     2401 tgatggtcgt catctacctg cctggacagc atggcctgca acgcgggcat cccgatgccg
     2461 ccggaagcga gaagaatcat aatggggaag gccatccagc ctcgcgtcgc gaacgccagc
     2521 aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa
     2581 cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc
     2641 gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc
     2701 cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg
     2761 gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag
     2821 ggcatcggtc ggcgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg
     2881 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gtaaggagat ggcgcccaac
     2941 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     3001 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     3061 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agagaatcca caggacgggt
     3121 gtggtcgcca tgatcgcgta gtcgatagtg gctccaagta gcgaagcgag caggactggg
     3181 cggcggccaa agcggtcgga cagtgctccg agaacgggtg cgcatagaaa ttgcatcaac
     3241 gcatatagcg ctagcagcac gccatagtga ctggcgatgc tgtcggaatg gacgatatcc
     3301 cgcaagaggc ccggcagtac cggcataacc aagcctatgc ctacagcatc cagggtgacg
     3361 gtgccgagga tgacgatgag cgcattgtta gatttcatac acggtgcctg actgcgttag
     3421 caatttaact gtgataaact accgcattaa agctaatcga tgataagctg tcaaacatga
     3481 gaattaattc atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3541 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3601 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     3661 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     3721 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     3781 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     3841 ttacgccaag ctatttaggt gacactatag aatactcaag cttgcatgac aggagctctc
     3901 ccgccattca gagaagaaac caattgtcca tattgcatca gacattgccg tcactgcgtc
     3961 ttttactggc tcttctcgct aacccaaccg gtaaccccgc ttattaaaag cattctgtaa
     4021 caaagcggga ccaaagccat gacaaaaacg cgtaacaaaa gtgtctataa tcacggcaga
     4081 aaagtccaca ttgattattt gcacggcgtc acactttgct atgccatagc atttttatcc
     4141 ataagattag cggatcctac ctgacgcttt ttatcgcaac tctctactgt ttctccatac
     4201 ccgttttagg aggtactgag catgcgtcta gagctcgagg tagatctgaa agcttgaatc
     4261 gatgtacgta ctataccggt gcgaaacggg aaggtaaaaa gacaaaaagt tgtttttaat
     4321 acctttaagt gataccagat ggcattgcgc catctggcag agtgattaac taaacatcgc
     4381 agtaatcgag gcgcttgcca gagagtggaa atgaacgtta aacccgacca tcgcgccgct
     4441 ggcaccttca tcgacatcaa tacgttctat atccagcgcg tgaacggtaa aaatgtagcg
     4501 atgagtttcg cctttcggcg gtgctgcgcc atcgtacccg gttttaccaa agtcggtacg
     4561 cgtctgcaaa acgccgtctg gcattgctac cagaccagag ccaaaccctt gcggtaatac
     4621 gcgggtatca gcgggtaagt taacaactac ccagtgccac cagccggagc cggttggcgc
     4681 atccgggtcg tagcaggtga caacaaaact tttcgttccc gcaggaacat catcccacgc
     4741 cagatgcggt gaaatattat cgccatcgta acccatgccg ttaaagacat gacgatgcgg
     4801 caatttatcg ccatcgcgca gatcgttact gatgagtttc atgaaccctc ctttcttgtt
     4861 tgcagaaagt gtagccagaa accctcacgc ggacttctcg ttattggcaa aaaaatgttt
     4921 catcctgtac cgcgcggtta accgctgcgg tcagacgctg caactgttgc gggagaataa
     4981 tatagggcgg catcaggtaa atcagtttgc caaaaggccg gatccagaca ccctgttcga
     5041 caaagaattt ttgcagcgcc gccatattca ccggatgagt ggtttcgacc acgccaatgg
     5101 cccccagtac gcgcacatcg gcaaccattt cggcatcacg ggcgggggca agttgctcgc
     5161 gcagctgtac ttcaatatcc gccacctgtt gctgccagtc gccagattcg agaatcgcca
     5221 ggctggcgtt tgctgccgcg caggccagcg gattgcccat aaaagttggc ccatgcataa
     5281 agcaaccggc ttcaccgtta ctgatggttt ctgcaacctc gcgcgtggtg agtgtggcgg
     5341 aaagggtcat tgtgccgccg gttaaggctt taccgaggca caaaatgtcc ggcgcgattt
     5401 ctgcatgttc acaggcaaac agtttcccgg tacgaccaaa tccagtggcg atctcgtcgg
     5461 caatcagcaa gataccttcg cgatcgcata ttttgcggat tcgttttaac cattccggat
     5521 ggtacatgcg catcccgcct gcgccctgga caatcggctc aatgatcacc gccgcgattt
     5581 catgacgatg cgccgccatc aggcgggcaa agcccaccat atcgcgctca tcccattcgc
     5641 catccatgcg gctttgcggg gcgggagcaa acaggttttc tggcaggtag cctttccaca
     5701 gactgtgcat tgagttatcc ggatcgcaca ccgacatcgc gccaaaggta tcgccatgat
     5761 aaccattgcg gaaggtcaga aaacgctggc gcgcttcgcc tttggcttgc cagtactgca
     5821 acgccatttt catcgccact tccaccgcta cggaaccgga gtccgcgaga aaaacgcact
     5881 ccagcggttg cggcgtcatc gccaccagtt tgcggcacag ctcaatggct ggcgcatggg
     5941 tgataccgcc aaacatcaca tgcgacatgg catcaatttg cgacttcatc gccgcattaa
     6001 gctgcgggtg attgtagccg tggatcgccg cccaccagga cgacataccg tcaaccaggc
     6061 gtctgccgtc agacaaaatc agctcgcaac cttcggcgct caccaccgga taaaccggca
     6121 gaggggaggt catggatgtg tatgggtgcc agatatggcg ttggtcaaag gcaagatcgt
     6181 ccgttgtcat aatcgacttg taaaccaaat tgaaaagatt taggtttaca agtctacacc
     6241 gaattaacaa caaaaaacac gttttggaga agccccatgg ctcaccgccc acgctggaca
     6301 ttgtcgcaag tcacagaatt atttgaaaaa ccgttgctgg atctgctgtt tgaagcgcag
     6361 caggtgcatc gccagcattt cgatcctcgt caggtgcagg tcagcacgtt gctgcatatg
     6421 gcgcgcctag gccaattggg cccgggcata tgggtgtgaa ataccgcaca gatgcgtaag
     6481 gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6541 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6601 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6661 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6721 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6781 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6841 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6901 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6961 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7021 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7081 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     7141 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7201 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7261 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7321 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7381 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     7441 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     7501 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7561 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7621 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7681 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7741 caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7801 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7861 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7921 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7981 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     8041 ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt
     8101 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     8161 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8221 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     8281 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     8341 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     8401 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     8461 gacattaacc tataaaaata ggcgtatcac gag
//
