LOCUS       sequence-320160-        8510 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  (parent plasmid pCFB257) pX - 3 -LoxP -KlLEU2 -pTEF1 - XylR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8510)
  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 8510)
  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_85
FEATURES             Location/Qualifiers
     source          1..8510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(15..180)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     101..125
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        229..635
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             666..686
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    1989..2022
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4313..4346
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(4389..4408)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4390..4408)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4418..4434)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4427..4449)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     protein_bind    complement(5097..5130)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(5600..5616)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5715..5732)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5886..6474)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5966..5985)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6596..7387,7388..7456))
                     /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     7219..7238
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7457..7548)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7590..7609
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7615..7634
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    7841..7874
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
ORIGIN
        1 ggaatgcgtg cgatgagcga cctcatgcta tacctgagaa agcaacctga cctacaggaa
       61 agagttactc aagaataaga attttcgttt taaaacctaa gagtcacttt aaaatttgta
      121 tacacttatt ttttttataa cttatttaat aataaaaatc ataaatcata agaaattcgc
      181 ttatttagaa gtgtcaacaa cgtatctacc aacggaatac gtgcgatgca cacaccatag
      241 cttcaaaatg tttctactcc ttttttactc ttccagattt tctcggactc cgcgcatcgc
      301 cgtaccactt caaaacaccc aagcacagca tactaaattt cccctctttc ttcctctagg
      361 gtgtcgttaa ttacccgtac taaaggtttg gaaaagaaaa aagagaccgc ctcgtttctt
      421 tttcttcgtc gaaaaaggca ataaaaattt ttatcacgtt tctttttctt gaaaattttt
      481 ttttttgatt tttttctctt tcgatgacct cccattgata tttaagttaa taaacggtct
      541 tcaatttctc aagtttcagt ttcatttttc ttgttctatt acaacttttt ttacttcttg
      601 ctcattagaa agaaagcata gcaatctaat ctaagtttta attacaaatc tgtcataaaa
      661 caatgcccaa gaagaaaagg aaagttaatc aaccagtaga aagacagcgt aggagaacta
      721 ctcaaagtgc tacaattcgt gacgtagctg caagagcagg tgtctctcct atgacagtct
      781 cacgtgtaat caatagagag tccacagtta aagaggaaac tagacagttg gttgaaaagg
      841 caatagccga ccttaactat gctcctaatc ctgcagccag atctttggca ggtagtgccc
      901 cttttagaat tggcttactg tacgataatc cttcaactgg ctacctttct gaatttctag
      961 ttggtgcctt agatgaatca agtagaaccg gtgctcaaat tgttatcgag aaatgtgctg
     1021 aaccagaatt agccagagct acacttgcta gattgttgaa aactggagtt gatggactta
     1081 tcttacctcc accattatgc gaatctccag aagttctggc cgagataaga gccgcaggag
     1141 ctgccgctgt cgcagtggca cctggtacag cttctgccga catggctact attagaatcg
     1201 acaacgaagc agctgcattt gagttgaccc agcatttgat tggcttgggt cacaaaagat
     1261 tcggattcat taagggtcat ccaaatcaaa ccgtgtctca acaaaggctt gatgggttta
     1321 tgactgctct taaggctgca gggatcccac aagagaatat cagagtggaa caaggttact
     1381 tcacatatcg ttcaggtcta gaagctgcag agagactact agcagccgaa cctaggccaa
     1441 ctgccatctt cgctgctaac gatgatatgg cagctgcaac agcaggcgta gcacatagac
     1501 taggcttgga tgtaccaggc gacgtgtcta tagtgggatt tgatgatact tccatagctg
     1561 ataacatttg gccaccatta acaacagttc accaaccaat tgccgctatg gccagagccg
     1621 ctgttgatct ggttctagaa gagatcagaa ggcatagaga tggtggtggc gaacctagac
     1681 aattgatgca tccacacact ctgatcgtta gagactcctc aggccctgct ggagtctaaa
     1741 tcgcgtgtat tcatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt
     1801 ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat ttcaaatttt
     1861 tctttttttt ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga
     1921 gaaggttttg ggacgctcga gatcgcgtca gctgaagctt cgtacgctgc aggtcgacaa
     1981 cccttaatat aacttcgtat aatgtatgct atacgaagtt attaggtcta gagatccgca
     2041 ggctaaccgg aacctgtatt atttagttta tgctacgtta aataaagacc tttcgttcac
     2101 ataactgaat gtgtaatggc cttgagattt caagcatacc aagttggtgg agacggggtc
     2161 gttacaaaag actctttaag ccaagatttc cttgacagcc ttggcgatag catcgccaac
     2221 ctcagtggta gagttagaac caccaaggtc accggttctg acacctgcat ccaagacatt
     2281 tctaacagct tcttcaagag ccctaccttc ttcaaccaaa tccaaggata acttcaacat
     2341 catagctgca gataagatgg tagcaattgg gttaaccttg tttgctggta aatctggggc
     2401 agaaccatga catggttcgt acaaaccgaa tgccttgtta gtgtcaggta gggaagctag
     2461 agatgcagaa ggtaataaac ccaaagaacc tggaataaca gaggcttcat cggagataat
     2521 atcaccaaac atgttgttgg taataacaac accgtttagc ttagttggtg atttaaccaa
     2581 aatcatagca gcagagtcga tcaattggtg ctgaacagtt aattgtggga actcagtctt
     2641 gatggtttct tcaacagtct ttctccacaa tctggaagag gcaagcacgt tagccttgtc
     2701 aagagaccag attggtaatg gtgggttttg ttgcaatgcc aagaaagcag ccattcttgt
     2761 aattctttga acttcaggaa cactgtattt ctcagagtcc caagcaactc cgtcaccttc
     2821 atcttctttt ctttcaccaa agtagatacc accaaccaat tctctaacga cgacgaaatc
     2881 ggtacccttt gcatattcag gcttcaaagg agaaagatct agtaaagaat cagaagcaaa
     2941 gttacatggt ctcaagttgg cgtatagacc caattccttt ctgatcttca atagaccttg
     3001 ttctggtcta actgcgcccg taccccattt tggaccacca acagcaccta gtaagacagc
     3061 atcggctttc ttagaggctt ctagagcttc atctggtaaa ggagtgccag tggcatcgat
     3121 ggcagcaccc ccgatcaagt gatgttggaa attgaactta atttctggac ggacttcagc
     3181 aatggcattc aagaccttaa tagcttcgtc agtaacttct ttaccgacgt gatcacccgg
     3241 taggacaacg atattcttag acatggtata atctgtgtag tgtgggatac tttttacttc
     3301 ttcaaatagg tatcaacaat aaaaataaat cgaagcaaat gtaggaatgc gttaaagcag
     3361 atgtacttac ttagagtaca tagatatatt tatataattc aatatataaa agtatatgaa
     3421 gcatctgatg ttgaacctgt catgactcta aactggaatg ggtagttatg cttctgaagg
     3481 ttttccgtta gaaaatgagt gcagggatca ggcccggaac cggttttagc ctgaaaggaa
     3541 aagaaaaaaa tgcggtgatt actagcacgt gactgcgctg aattggaatc taccacaagt
     3601 acagcgaatg gcaagaggga aggggaatat attgtgcctg gcggtagcgt ctgctatatc
     3661 aaatgttaaa cataactaaa tgcagtagtg aatagagtaa tgtatttcta catatgtggg
     3721 tagggccaat gggagcccga tgtctattgt acagatattt tccttttata gttaatcaat
     3781 tcaacttgga aatcattgag tcgatatcta cttggataac tggagccgtg gcagcgtttg
     3841 aaatttgacg agtcttcttg ttttgttcgc gcattttcag tttattcaaa cgggagatgg
     3901 caccgaatcc actgacttct ctgttcaatt tgaatggtaa taggaagaaa gtacatttcg
     3961 agaaattttc gcccaacgtc gaaatctcat ggatgatatc ctcgatacaa atgacacctt
     4021 catcacctaa tttctcttca acgatattgt tgtcattcaa gattatctct tttggttctt
     4081 tatcttctgg cctttgccac attattctag atctcttttg gattaaagaa cggatagatg
     4141 caagtgaagg ttgtccgata acaatatatg gtgatacaag tttcaacaat gggtacacgt
     4201 ccttggtaag tttgatgaaa acaccagtgt tcaactcaac aagtctcaaa agagataaca
     4261 ctttgtaagc ctttgctggg atcttcacag cgagctctcg agaaccctta atataacttc
     4321 gtataatgta tgctatacga agttattagg tgatatcaga tccactagtg gcctatgcac
     4381 ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc
     4441 gtgactggga aaaccctggc gttacccctg caggactagt gctgaggcat taattgtgtc
     4501 cgcgtttcta aggccacttt tcaatgaaac ggatattgat atgctagtaa aaggacgagc
     4561 tcaagagcga aaatataagt aaagaattcg agtgcacttg tctccatgca gcaagatttc
     4621 atatgagtct tttttatctt tttacttttt acattacacg atatgcactt tatgaaaatt
     4681 taacgaggtt ggaagccgga taatcaacca aaatcaggca cgaaggcaca ctcgtatatg
     4741 catgttgttg aaactctgtt acgctgaact aacaatcaca catgtagagg tcaccgggaa
     4801 aagttgcgac cccatggaag gtcgatctct tcgtttggct ttgcttggct ggcggcattg
     4861 cgcttcttcg cttatacccg tctcttgacg ctcgagctcg ttcattgaga tacctttatt
     4921 cttgcacatt ttctggcttt tttcgctact cgggtacatg taatcatgca cacagaaggt
     4981 gctgtagggt gaaagttcct ttgtgctgtc gtttgttttt aatgccaaac tttccggtga
     5041 tcaataacca cctcattaag acctcagcgc ggccgcaaat ttaaataaaa tgaagtgaag
     5101 ttcctatact ttctagagaa taggaacttc tatagtgagt cgaataaggg cgacacaaaa
     5161 tttattctaa atgcataata aatactgata acatcttata gtttgtatta tattttgtat
     5221 tatcgttgac atgtataatt ttgatatcaa aaactgattt tccctttatt attttcgaga
     5281 tttattttct taattctctt taacaaacta gaaatattgt atatacaaaa aatcataaat
     5341 aatagatgaa tagtttaatt ataggtgttc atcaatcgaa aaagcaacgt atcttattta
     5401 aagtgcgttg cttttttctc atttataagg ttaaataatt ctcatatatc aagcaaagtg
     5461 acaggcgccc ttaaatattc tgacaaatgc tctttcccta aactcccccc ataaaaaaac
     5521 ccgccgaagc gggtttttac gttatttgcg gattaacgat tactcgttat cagaaccgcc
     5581 cagggggccc gagcttaaga ctggccgtcg ttttacaaca cagaaagagt ttgtagaaac
     5641 gcaaaaaggc catccgtcag gggccttctg cttagtttga tgcctggcag ttccctactc
     5701 tcgccttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5761 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5821 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5881 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5941 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     6001 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     6061 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     6121 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     6181 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     6241 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6301 tgggctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     6361 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6421 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6481 cctttgatct tttctacggg gtctgacgct cagtggaacg acgcgcgcgt aactcacgtt
     6541 aagggatttt ggtcatgagc ttgcgccgtc ccgtcaagtc agcgtaatgc tctgcttacc
     6601 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6661 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagcg
     6721 ctgcgatgat accgcgagaa ccacgctcac cggctccgga tttatcagca ataaaccagc
     6781 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6841 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6901 ttgccatcgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6961 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7021 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7081 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7141 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7201 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7261 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7321 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7381 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7441 aatgttgaat actcatattc ttcctttttc aatattattg aagcatttat cagggttatt
     7501 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggtcagtg
     7561 ttacaaccaa ttaaccaatt ctgaacatta tcgcgagccc atttatacct gaatatggct
     7621 cataacaccc cttgtttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
     7681 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggactcccca tgcgagagta
     7741 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgccc
     7801 gggctaatta tggggtgtcg cccttattcg actctatagt gaagttccta ttctctagaa
     7861 agtataggaa cttctgaagt ggggatttaa atgcggccgc gctgagggtt taatcgagat
     7921 ctttgtgttc ggttacccgg ctcagatcct aacttcttct tttggtatgt ttattcgtat
     7981 aagttactgt tgtccacagg caatactctg cagaaaatta aaacggcatt aatgctagga
     8041 caaccagaat tgttactact gtatgtgcga tagttgataa ctgcaacatt atgcccggta
     8101 tattctcaaa aaaccctatt actgcatacg aagaaatcgc aagagaaatc tttcggtttg
     8161 gaaaagctca ctgtgaggtt ccttggagcc aatagtaata cagcacaatc caaggaaaaa
     8221 tctggcctat atgcaaggaa ggagagatag tcaaaagcat tctttcccct agaagttggt
     8281 gcatatatgg catcgttaaa acatattacc cccaaaattt cttctctaaa cgatgtgctt
     8341 ggcctttgtt ttggtttttg atgtcggtcg tttgaggccc cttgcggaaa atcgagatcg
     8401 ccgaatggca cgcgagggaa gggaaataag gtttaaaggc actgaaacaa taggcaagaa
     8461 gtaggcgaga gccgacatac gagactatta agtcctcagc gagctcgcat
//
