LOCUS       sequence-320170-        7325 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pXI-3-LoxP-KlURA3-pTEF1- FapO_94-yeGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7325)
  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 7325)
  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: pFapO_94
FEATURES             Location/Qualifiers
     source          1..7325
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(153..186)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      complement(234..423)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             906..1622
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1670..1835
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1725..1749)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    complement(2489..2522)
                     /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(2729..2748)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(2754..2773)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        2815..2906
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2907..2975,2976..3767)
                     /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     complement(3125..3144)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3889..4477
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4378..4397
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4631..4648
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4747..4763
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5233..5266
                     /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     5890..5912
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5905..5921
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5931..5950
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5931..5949
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(5993..6026)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 aatcagcgct ccccattaat tatacaggaa acttaataga acaaatcaca tatttaatct
       61 aatagccacc tgcattggca cggtgcaaca ctacttcaac ttcatcttac aaaaagatca
      121 cgtgatctgt tgtattggga tctctagacc taataacttc gtatagcata cattatacga
      181 agttatatta agggttgtcg acctgcagcg tacgaagctt cagctgacgc gatcttcgag
      241 cgtcccaaaa ccttctcaag caaggttttc agtataatgt tacatgcgta cacgcgtctg
      301 tacagaaaaa aaagaaaaat ttgaaatata aataacgttc ttaatactaa cataactata
      361 aaaaaataaa tagggaccta gacttcaggt tgtctaactc cttccttttc ggttagagca
      421 gatgaataca cgcgatgcac acaccatagc ttcaaaatgt ttctactcct tttttactct
      481 tccagatttt ctcggactcc gcgcatcgcc gtaccacttc aaaacaccca agcacagcat
      541 actaaatttc ccctctttct tcctctaggg tgtcgttaat tacccgtact aaaggtttgg
      601 aaaagaaaaa agagaccgcc tcgtttcttt ttcttcgtcg aaaaaggcaa taaaaatttt
      661 tatcacgttt ctttttcttg aaaatttttt tttttgattt ttttctcttt cgatgacctc
      721 ccattgatat ttaagttaat aaacggtctt caatttctca agaattatat actactatta
      781 gtacctagtc ttaatttttc agtttcattt ttcttgttct attacaactt tttttacttc
      841 ttgctcatta gaaagaaagc atagcaatct aatctaagtt ttaattacaa agtgcaggta
      901 aaacaatgtc taaaggtgaa gaattattca ctggtgttgt cccaattttg gttgaattag
      961 atggtgatgt taatggtcac aaattttctg tctccggtga aggtgaaggt gatgctactt
     1021 acggtaaatt gaccttaaaa tttatttgta ctactggtaa attgccagtt ccatggccaa
     1081 ccttagtcac tactttcggt tatggtgttc aatgttttgc gagataccca gatcatatga
     1141 aacaacatga ctttttcaag tctgccatgc cagaaggtta tgttcaagaa agaactattt
     1201 ttttcaaaga tgacggtaac tacaagacca gagctgaagt caagtttgaa ggtgatacct
     1261 tagttaatag aatcgaatta aaaggtattg attttaaaga agatggtaac attttaggtc
     1321 acaaattgga atacaactat aactctcaca atgtttacat catggctgac aaacaaaaga
     1381 atggtatcaa agttaacttc aaaattagac acaacattga agatggttct gttcaattag
     1441 ctgaccatta tcaacaaaat actccaattg gtgatggtcc agtcttgtta ccagacaacc
     1501 attacttatc cactcaatct gccttatcca aagatccaaa cgaaaagaga gaccacatgg
     1561 tcttgttaga atttgttact gctgctggta ttacccatgg tatggatgaa ttgtacaaat
     1621 aaatcgcacg tattccgttg gtagatacgt tgttgacact tctaaataag cgaatttctt
     1681 atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta
     1741 aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt tcctgtaggt
     1801 caggttgctt tctcaggtat agcatgaggt cgctcatcgc acgcattcca tgcgagctcg
     1861 ctgaggactt aataatcaga cgcacgcttg gcgtcaatat cctaaaacgg aaaactaatc
     1921 agttagatgt gtctttttta ggtaaaaaaa ggcaaccaat tttttgctgg ctagcgatgc
     1981 gacaataaac gcgcgcagcg aataaaaata agatggcgga ggagcagaga cttcttttct
     2041 tagaagaggt tttgttaccg tatgggggta tgatggttag atggctgaat aaaaacgtta
     2101 attaaaggaa tacagaagag gaagcgcccc gatttctttt ctgttccaca tgaaagcagc
     2161 cggttgggtg gtagatctct aatgacatcc aaactacaaa accgagattg gacatatagc
     2221 acactctgca tttctacttt cctttctccc tacagtttgc tcgatctgaa ccgatgcttc
     2281 tttgattgga caagatgata gaccatgact ccctgtatgt attgcagaga ttgccactac
     2341 taaccaagaa aagttgaacg tgtcgtacgt ataaaacgat ttatgcttag gctttactga
     2401 aaaaaaaaaa gtaaaaagag gatgcgcaaa cgcatagagc aagtaactat taaaccctca
     2461 gcgcggccgc atttaaatcc ccacttcaga agttcctata ctttctagag aataggaact
     2521 tcactataga gtcgaataag ggcgacaccc cataattagc ccgggcgaaa ggcccagtct
     2581 ttcgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg catggggagt
     2641 ccccacacta ccatcggcgc tacggcgttt cacttctgag ttcggcatgg ggtcaggtgg
     2701 gaccaccgcg ctactgccgc caggcaaaca aggggtgtta tgagccatat tcaggtataa
     2761 atgggctcgc gataatgttc agaattggtt aattggttgt aacactgacc cctatttgtt
     2821 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     2881 ttcaataata ttgaaaaagg aagaatatga gtattcaaca tttccgtgtc gcccttattc
     2941 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3001 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     3061 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     3121 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     3181 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     3241 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3301 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     3361 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     3421 caaacgacga gcgtgacacc acgatgcctg tagcgatggc aacaacgttg cgcaaactat
     3481 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     3541 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3601 aatccggagc cggtgagcgt ggttctcgcg gtatcatcgc agcgctgggg ccagatggta
     3661 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     3721 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaagca gagcattacg
     3781 ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag ttacgcgcgc
     3841 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3901 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3961 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4021 gataccaaat actgttcttc tagtgtagcc gtagttagcc caccacttca agaactctgt
     4081 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4141 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4201 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4261 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4321 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4381 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4441 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4501 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     4561 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     4621 gaccgagcgc agcgagtcag tgagcgagga agcggaaggc gagagtaggg aactgccagg
     4681 catcaaacta agcagaaggc ccctgacgga tggccttttt gcgtttctac aaactctttc
     4741 tgtgttgtaa aacgacggcc agtcttaagc tcgggccccc tgggcggttc tgataacgag
     4801 taatcgttaa tccgcaaata acgtaaaaac ccgcttcggc gggttttttt atggggggag
     4861 tttagggaaa gagcatttgt cagaatattt aagggcgcct gtcactttgc ttgatatatg
     4921 agaattattt aaccttataa atgagaaaaa agcaacgcac tttaaataag atacgttgct
     4981 ttttcgattg atgaacacct ataattaaac tattcatcta ttatttatga ttttttgtat
     5041 atacaatatt tctagtttgt taaagagaat taagaaaata aatctcgaaa ataataaagg
     5101 gaaaatcagt ttttgatatc aaaattatac atgtcaacga taatacaaaa tataatacaa
     5161 actataagat gttatcagta tttattatgc atttagaata aattttgtgt cgcccttatt
     5221 cgactcacta tagaagttcc tattctctag aaagtatagg aacttcactt cattttattt
     5281 aaatttgcgg ccgcgctgag gtcttaattg agaatccgga ccagcagata atgcatgcag
     5341 tggtgttttg atgtataagc ttgacatttt tttgcttttc ccgttatgag aaatgttttt
     5401 tttagactga aaataaaaga atggatagta ttaaaatagt aatgaacgta ataaggatag
     5461 gacaggagcg ataggaaaag atagaatttt ggagcttttt ttccctcttt ttttgagcag
     5521 gagccaatag tagtcggaaa gatgagtgca tagatttttc gtattttatt tcgagtaaaa
     5581 atatcacgct cttaggttca ataaaagaat atactatttc ttcaaaatgc ttctctaatc
     5641 tattttttta ttttccttaa taactgaaat attatttttt gatgtaaaat gagtgaacaa
     5701 gaataaggga tttttggtat gtttgcaagt tctgtggagt tccatgtata aggtatacat
     5761 attagaaaag ctttacgatt atatactaac atcatgtaca aaacagttta ataatgatct
     5821 gtattgctgg ctcaatccac gtaaattaat gcctcagcac tagtcctgca ggggtaacgc
     5881 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg taatacgact
     5941 cactataggg cgaattgggt gcataggcca ctagtggatc tgatatcacc taataacttc
     6001 gtatagcata cattatacga agttatatta agggttctcg agagctcgtt ttatttaggt
     6061 tctatcgagg agaaaaagcg acaagaagag atagaccatg gataaatgat tatgttctaa
     6121 acactcctca gaagctcatc gaactgtcat cctgcgtgaa gattaaaatc caacttagaa
     6181 atttcgagct tacggagaca atcatatggg agaagcaatt ggaagataga aaaaaggtac
     6241 tcggtacata aatatatgtg attctgggta gaagatcggt ctgcattgga tggtggtaac
     6301 gcattttttt acacacatta cttgcctcga gcatcaaatg gtggttattc gtggatctat
     6361 atcacgtgat ttgcttaaga attgtcgttc atggtgacac ttttagcttt gacatgatta
     6421 agctcatctc aattgatgtt atctaaagtc atttcaacta tctaagatgt ggttgtgatt
     6481 gggccatttt gtgaaagcca gtacgccagc gtcaatacac tcccgtcaat tagttgcacc
     6541 atgtccacaa aatcatatac cagtagagct gagactcatg caagtccggt tgcatcgaaa
     6601 cttttacgtt taatggatga aaagaagacc aatttgtgtg cttctcttga cgttcgttcg
     6661 actgatgagc tattgaaact tgttgaaacg ttgggtccat acatttgcct tttgaaaaca
     6721 cacgttgata tcttggatga tttcagttat gagggtactg tcgttccatt gaaagcattg
     6781 gcagagaaat acaagttctt gatatttgag gacagaaaat tcgccgatat cggtaacaca
     6841 gtcaaattac aatatacatc gggcgtttac cgtatcgcag aatggtctga tatcaccaac
     6901 gcccacgggg ttactggtgc tggtattgtt gctggcttga aacaaggtgc gcaagaggtc
     6961 accaaagaac caaggggatt attgatgctt gctgaattgt cttccaaggg ttctctagca
     7021 cacggtgaat atactaaggg taccgttgat attgcaaaga gtgataaaga tttcgttatt
     7081 gggttcattg ctcagaacga tatgggagga agagaagaag ggtttgattg gctaatcatg
     7141 accccaggtg taggtttaga cgacaaaggc gatgcattgg gtcagcagta cagaaccgtc
     7201 gacgaagttg taagtggtgg atcagatatc atcattgttg gcagaggact tttcgccaag
     7261 ggtagagatc ctaaggttga aggtgaaaga tacagaaatg ctggatggga agcgtaccaa
     7321 aagag
//
