LOCUS       sequence-319278-        6373 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Wild type VanR from Caulobacter crescentus. The gene is under the 
            control of the TEF1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6373)
  AUTHORS   D'Ambrosio, V., Pramanik, S., Goroncy, K., Jakociunas, T., 
            Schonauer, D., Davari, H. D., Schwaneberg, U., Keasling, J.D., 
            Jensen, M.K.
  TITLE     Directed evolution of VanR biosensor specificity in yeast
  JOURNAL   Biotechnology Notes, Volume 1, 2020, Pages 9-15
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"; journalName: 
            "Biotechnology Notes, Volume 1, 2020, Pages 9-15"
COMMENT     Sequence Label: VanR-wt
FEATURES             Location/Qualifiers
     source          1..6373
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     32..56
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        160..566
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(1533..1553)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1533..1551)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1572..1588)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1572..1588)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1585..1607)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1596..1612
                     /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(1620..1626,1627..1644,1645..1650))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1665..1686
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1803..1820)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1974..2562)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2054..2073)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2733..3524,3525..3593))
                     /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     3356..3375
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3594..3698)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    3735..4238
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     4279..4297
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4335..4357)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4457..4476
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4500..4715
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4716..5519
                     /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(5650..6105)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5787..5808)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5999..6018
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6231..6253
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6245..6262
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6246..6262
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6272..6291
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6272..6290
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(join(6319..6373,1..111))
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
ORIGIN
        1 caagaataag aattttcgtt ttaaaaccta agagtcactt taaaatttgt atacacttat
       61 tttttttata acttatttaa taataaaaat cataaatcat aagaaattcg cttatttaga
      121 agtgtcaaca acgtatctac caacggaatg cgtgcgatgc acacaccata gcttcaaaat
      181 gtttctactc cttttttact cttccagatt ttctcggact ccgcgcatcg ccgtaccact
      241 tcaaaacacc caagcacagc atactaaatt tcccctcttt cttcctctag ggtgtcgtta
      301 attacccgta ctaaaggttt ggaaaagaaa aaagagaccg cctcgtttct ttttcttcgt
      361 cgaaaaaggc aataaaaatt tttatcacgt ttctttttct tgaaaatttt tttttttgat
      421 ttttttctct ttcgatgacc tcccattgat atttaagtta ataaacggtc ttcaatttct
      481 caagtttcag tttcattttt cttgttctat tacaactttt tttacttctt gctcattaga
      541 aagaaagcat agcaatctaa tctaagtttt aattacaaat ctgtcataaa acaatggaca
      601 tgccgagaat caaacccggt cagagggtta tgatggcgtt gcgtaagatg atcgctagtg
      661 gggaaatcaa atccggcgaa aggattgcag agatcccaac cgcagccgca ctgggggttt
      721 ccaggatgcc agtccgtaca gcattgcgta gtttggaaca agagggattg gtggttcgtt
      781 tgggcgccag aggttacgcg gctcgtggtg tcagcagcga ccaaataaga gacgcgatcg
      841 aagttagggg agtactggaa gggttcgctg ccaggcgttt agctgagcgt gggatgacag
      901 cggaaaccca tgcccgtttt gtagctctaa ttgctgaggg agaggcttta tttgcagcag
      961 ggcgtctaaa cggagaggat cttgacaggt atgctgcgta caaccaagca tttcacgaca
     1021 cccttgtgtc cgcggcaggc aatggcgcgg tagagtccgc ccttgcgcgt aacggcttcg
     1081 agcccttcgc cgctgctgga gcattagcac ttgacctaat ggacttgcca gcggagtatg
     1141 aacacctgct ggcggcccac agacagcatc aagccgtatt ggatgcggtg tcatgtggcg
     1201 atgcagaggg agcagagaga atcatgaggg accacgctct tgcggcgatc cgtaacgcaa
     1261 aagtttttga agctgctgct tccgctgggg ctccgctagg tgcggcctgg agtataagag
     1321 cagactgata atcgcgtgca ttcatccgct ctaaccgaaa aggaaggagt tagacaacct
     1381 gaagtctagg tccctattta tttttttata gttatgttag tattaagaac gttatttata
     1441 tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa
     1501 accttgcttg agaagagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg
     1561 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     1621 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     1681 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     1741 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     1801 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     1861 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     1921 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1981 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2041 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2101 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2161 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2221 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2281 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2341 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2401 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2461 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2521 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2581 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2641 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     2701 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     2761 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     2821 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     2881 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     2941 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3001 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     3061 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3121 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3181 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3241 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3301 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     3361 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     3421 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     3481 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     3541 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     3601 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     3661 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     3721 cctgggtcct tttcatcacg tgctataaaa ataattataa tttaaatttt ttaatataaa
     3781 tatataaatt aaaaatagaa agtaaaaaaa gaaattaaag aaaaaatagt ttttgttttc
     3841 cgaagatgta aaagactcta gggggatcgc caacaaatac taccttttat cttgctcttc
     3901 ctgctctcag gtattaatgc cgaattgttt catcttgtct gtgtagaaga ccacacacga
     3961 aaatcctgtg attttacatt ttacttatcg ttaatcgaat gtatatctat ttaatctgct
     4021 tttcttgtct aataaatata tatgtaaagt acgctttttg ttgaaatttt ttaaaccttt
     4081 gtttattttt ttttcttcat tccgtaactc ttctaccttc tttatttact ttctaaaatc
     4141 caaatacaaa acataaaaat aaataaacac agagtaaatt cccaaattat tccatcatta
     4201 aaagatacga ggcgcgtgta agttacaggc aagcgatccg tctaagaaac cattattatc
     4261 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     4321 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     4381 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     4441 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca taccaccttt
     4501 tcaattcatc attttttttt tattcttttt tttgatttcg gtttccttga aatttttttg
     4561 attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt agattggtat
     4621 atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta acccaactgc
     4681 acagaacaaa aacctgcagg aaacgaagat aaatcatgtc gaaagctaca tataaggaac
     4741 gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg cacgaaaagc
     4801 aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg gagttagttg
     4861 aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg actgattttt
     4921 ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat tttttactct
     4981 tcgaagacag aaaatttgct gacattggta atacagtcaa attgcagtac tctgcgggtg
     5041 tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg ggcccaggta
     5101 ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga ggccttttga
     5161 tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact aagggtactg
     5221 ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa agagacatgg
     5281 gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt ttagatgaca
     5341 agggagacgc attgggtcaa cagtatagaa ccgtggatga tgtggtctct acaggatctg
     5401 acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag gtagagggtg
     5461 aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag caaaactaaa
     5521 aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct tcaatttaat
     5581 tatatcagtt attaccctat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     5641 cgcatcagga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     5701 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     5761 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     5821 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     5881 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     5941 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     6001 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     6061 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt cgccattcag
     6121 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     6181 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     6241 acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattggg
     6301 taccgggccc cccctcgaga gcgacctcat gctatacctg agaaagcaac ctgacctaca
     6361 ggaaagagtt act
//
