LOCUS       sequence-319302-        6373 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTEF1-VanR inserted in pRS416 backbone .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6373)
  AUTHORS   Ambri F, D'Ambrosio V, Di Blasi R, Maury J, Jacobsen SAB, McCloskey 
            D, Jensen MK, Keasling JD
  TITLE     High-Resolution Scanning of Optimal Biosensor Reporter Promoters in 
            Yeast.
  JOURNAL   ACS Synth Biol. 2020 Feb 21;9(2):218-226. doi: 
            10.1021/acssynbio.9b00333. Epub 2020 Jan 31.
  PUBMED    31935067
REFERENCE   2  (bases 1 to 6373)
  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"; doi: 
            "10.1021/acssynbio.9b00333"; journalName: "ACS Synth Biol"; date: 
            "2020-02-21- 21"; volume: "9"; issue: "2"; pages: "218-226"
COMMENT     Sequence Label: pVD_01
FEATURES             Location/Qualifiers
     source          1..6373
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        100..506
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(1473..1493)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1473..1491)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1512..1528)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1512..1528)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1525..1547)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1536..1552
                     /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)."
     promoter        complement(join(1560..1566,1567..1584,1585..1590))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1605..1626
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1743..1760)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1914..2502)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1994..2013)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2673..3464,3465..3533))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3296..3315
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3534..3638)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    3675..4178
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     4219..4237
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4275..4297)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4397..4416
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4440..4655
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4656..5459
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(5590..6045)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5727..5748)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5939..5958
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6171..6193
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6185..6202
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6186..6202
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6212..6231
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6212..6230
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(join(6259..6373,1..51))
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     6345..6369
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
ORIGIN
        1 tttttttata acttatttaa taataaaaat cataaatcat aagaaattcg cttatttaga
       61 agtgtcaaca acgtatctac caacggaatg cgtgcgatgc acacaccata gcttcaaaat
      121 gtttctactc cttttttact cttccagatt ttctcggact ccgcgcatcg ccgtaccact
      181 tcaaaacacc caagcacagc atactaaatt tcccctcttt cttcctctag ggtgtcgtta
      241 attacccgta ctaaaggttt ggaaaagaaa aaagagaccg cctcgtttct ttttcttcgt
      301 cgaaaaaggc aataaaaatt tttatcacgt ttctttttct tgaaaatttt tttttttgat
      361 ttttttctct ttcgatgacc tcccattgat atttaagtta ataaacggtc ttcaatttct
      421 caagtttcag tttcattttt cttgttctat tacaactttt tttacttctt gctcattaga
      481 aagaaagcat agcaatctaa tctaagtttt aattacaaat ctgtcataaa acaatggaca
      541 tgccgagaat caaacccggt cagagggtta tgatggcgtt gcgtaagatg atcgctagtg
      601 gggaaatcaa atccggcgaa aggattgcag agatcccaac cgcagccgca ctgggggttt
      661 ccaggatgcc agtccgtaca gcattgcgta gtttggaaca agagggattg gtggttcgtt
      721 tgggcgccag aggttacgcg gctcgtggtg tcagcagcga ccaaataaga gacgcgatcg
      781 aagttagggg agtactggaa gggttcgctg ccaggcgttt agctgagcgt gggatgacag
      841 cggaaaccca tgcccgtttt gtagctctaa ttgctgaggg agaggcttta tttgcagcag
      901 ggcgtctaaa cggagaggat cttgacaggt atgctgcgta caaccaagca tttcacgaca
      961 cccttgtgtc cgcggcaggc aatggcgcgg tagagtccgc ccttgcgcgt aacggcttcg
     1021 agcccttcgc cgctgctgga gcattagcac ttgacctaat ggacttgcca gcggagtatg
     1081 aacacctgct ggcggcccac agacagcatc aagccgtatt ggatgcggtg tcatgtggcg
     1141 atgcagaggg agcagagaga atcatgaggg accacgctct tgcggcgatc cgtaacgcaa
     1201 aagtttttga agctgctgct tccgctgggg ctccgctagg tgcggcctgg agtataagag
     1261 cagactgata atcgcgtgca ttcatccgct ctaaccgaaa aggaaggagt tagacaacct
     1321 gaagtctagg tccctattta tttttttata gttatgttag tattaagaac gttatttata
     1381 tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa
     1441 accttgcttg agaagagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg
     1501 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     1561 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     1621 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     1681 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     1741 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     1801 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     1861 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1921 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     1981 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2041 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2101 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2161 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2221 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2281 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2341 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2401 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2461 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2521 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2581 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     2641 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     2701 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     2761 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     2821 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     2881 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     2941 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     3001 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3061 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3121 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3181 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3241 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     3301 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     3361 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     3421 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     3481 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     3541 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     3601 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     3661 cctgggtcct tttcatcacg tgctataaaa ataattataa tttaaatttt ttaatataaa
     3721 tatataaatt aaaaatagaa agtaaaaaaa gaaattaaag aaaaaatagt ttttgttttc
     3781 cgaagatgta aaagactcta gggggatcgc caacaaatac taccttttat cttgctcttc
     3841 ctgctctcag gtattaatgc cgaattgttt catcttgtct gtgtagaaga ccacacacga
     3901 aaatcctgtg attttacatt ttacttatcg ttaatcgaat gtatatctat ttaatctgct
     3961 tttcttgtct aataaatata tatgtaaagt acgctttttg ttgaaatttt ttaaaccttt
     4021 gtttattttt ttttcttcat tccgtaactc ttctaccttc tttatttact ttctaaaatc
     4081 caaatacaaa acataaaaat aaataaacac agagtaaatt cccaaattat tccatcatta
     4141 aaagatacga ggcgcgtgta agttacaggc aagcgatccg tctaagaaac cattattatc
     4201 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     4261 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     4321 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     4381 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca taccaccttt
     4441 tcaattcatc attttttttt tattcttttt tttgatttcg gtttccttga aatttttttg
     4501 attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt agattggtat
     4561 atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta acccaactgc
     4621 acagaacaaa aacctgcagg aaacgaagat aaatcatgtc gaaagctaca tataaggaac
     4681 gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg cacgaaaagc
     4741 aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg gagttagttg
     4801 aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg actgattttt
     4861 ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat tttttactct
     4921 tcgaagacag aaaatttgct gacattggta atacagtcaa attgcagtac tctgcgggtg
     4981 tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg ggcccaggta
     5041 ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga ggccttttga
     5101 tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact aagggtactg
     5161 ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa agagacatgg
     5221 gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt ttagatgaca
     5281 agggagacgc attgggtcaa cagtatagaa ccgtggatga tgtggtctct acaggatctg
     5341 acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag gtagagggtg
     5401 aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag caaaactaaa
     5461 aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct tcaatttaat
     5521 tatatcagtt attaccctat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     5581 cgcatcagga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     5641 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     5701 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     5761 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     5821 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     5881 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     5941 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     6001 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt cgccattcag
     6061 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     6121 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     6181 acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattggg
     6241 taccgggccc cccctcgaga gcgacctcat gctatacctg agaaagcaac ctgacctaca
     6301 ggaaagagtt actcaagaat aagaattttc gttttaaaac ctaagagtca ctttaaaatt
     6361 tgtatacact tat
//
