LOCUS       sequence-320166-        7308 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pXI - 3 -LoxP -KlURA3 -pTEF1 - TetO_105 -yeGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7308)
  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 7308)
  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: pTetO_105
FEATURES             Location/Qualifiers
     source          1..7308
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(163..196)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      complement(244..433)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             899..1615
                     /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      1663..1828
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1718..1742)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    complement(2482..2515)
                     /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(2722..2741)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(2747..2766)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        2808..2899
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2900..2968,2969..3760)
                     /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(3118..3137)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3882..4470
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4371..4390
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4624..4641
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4740..4756
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5226..5259
                     /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     5883..5905
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5898..5914
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5924..5943
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5924..5942
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(5986..6019)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 accaaaagag aatcagcgct ccccattaat tatacaggaa acttaataga acaaatcaca
       61 tatttaatct aatagccacc tgcattggca cggtgcaaca ctacttcaac ttcatcttac
      121 aaaaagatca cgtgatctgt tgtattggga tctctagacc taataacttc gtatagcata
      181 cattatacga agttatatta agggttgtcg acctgcagcg tacgaagctt cagctgacgc
      241 gatcttcgag cgtcccaaaa ccttctcaag caaggttttc agtataatgt tacatgcgta
      301 cacgcgtctg tacagaaaaa aaagaaaaat ttgaaatata aataacgttc ttaatactaa
      361 cataactata aaaaaataaa tagggaccta gacttcaggt tgtctaactc cttccttttc
      421 ggttagagcg gatgaataca cgcgatgcac acaccatagc ttcaaaatgt ttctactcct
      481 tttttactct tccagatttt ctcggactcc gcgcatcgcc gtaccacttc aaaacaccca
      541 agcacagcat actaaatttc ccctctttct tcctctaggg tgtcgttaat tacccgtact
      601 aaaggtttgg aaaagaaaaa agagaccgcc tcgtttcttt ttcttcgtcg aaaaaggcaa
      661 taaaaatttt tatcacgttt ctttttcttg aaaatttttt tttttgattt ttttctcttt
      721 cgatgacctc ccattgatat ttaagttaat aaacggtctt cctctatcat tgatagagaa
      781 tttctcaagt ttcagtttca tttttcttgt tctattacaa ctttttttac ttcttgctca
      841 ttagaaagaa agcatagcaa tctaatctaa gttttaatta caaagtgcag gtaaaacaat
      901 gtctaaaggt gaagaattat tcactggtgt tgtcccaatt ttggttgaat tagatggtga
      961 tgttaatggt cacaaatttt ctgtctccgg tgaaggtgaa ggtgatgcta cttacggtaa
     1021 attgacctta aaatttattt gtactactgg taaattgcca gttccatggc caaccttagt
     1081 cactactttc ggttatggtg ttcaatgttt tgcgagatac ccagatcata tgaaacaaca
     1141 tgactttttc aagtctgcca tgccagaagg ttatgttcaa gaaagaacta tttttttcaa
     1201 agatgacggt aactacaaga ccagagctga agtcaagttt gaaggtgata ccttagttaa
     1261 tagaatcgaa ttaaaaggta ttgattttaa agaagatggt aacattttag gtcacaaatt
     1321 ggaatacaac tataactctc acaatgttta catcatggct gacaaacaaa agaatggtat
     1381 caaagttaac ttcaaaatta gacacaacat tgaagatggt tctgttcaat tagctgacca
     1441 ttatcaacaa aatactccaa ttggtgatgg tccagtcttg ttaccagaca accattactt
     1501 atccactcaa tctgccttat ccaaagatcc aaacgaaaag agagaccaca tggtcttgtt
     1561 agaatttgtt actgctgctg gtattaccca tggtatggat gaattgtaca aataaatcgc
     1621 acgtattccg ttggtagata cgttgttgac acttctaaat aagcgaattt cttatgattt
     1681 atgattttta ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac
     1741 tcttaggttt taaaacgaaa attcttattc ttgagtaact ctttcctgta ggtcaggttg
     1801 ctttctcagg tatagcatga ggtcgctcat cgcacgcatt ccatgcgagc tcgctgagga
     1861 cttaataatc agacgcacgc ttggcgtcaa tatcctaaaa cggaaaacta atcagttaga
     1921 tgtgtctttt ttaggtaaaa aaaggcaacc aattttttgc tggctagcga tgcgacaata
     1981 aacgcgcgca gcgaataaaa ataagatggc ggaggagcag agacttcttt tcttagaaga
     2041 ggttttgtta ccgtatgggg gtatgatggt tagatggctg aataaaaacg ttaattaaag
     2101 gaatacagaa gaggaagcgc cccgatttct tttctgttcc acatgaaagc agccggttgg
     2161 gtggtagatc tctaatgaca tccaaactac aaaaccgaga ttggacatat agcacactct
     2221 gcatttctac tttcctttct ccctacagtt tgctcgatct gaaccgatgc ttctttgatt
     2281 ggacaagatg atagaccatg actccctgta tgtattgcag agattgccac tactaaccaa
     2341 gaaaagttga acgtgtcgta cgtataaaac gatttatgct taggctttac tgaaaaaaaa
     2401 aaagtaaaaa gaggatgcgc aaacgcatag agcaagtaac tattaaaccc tcagcgcggc
     2461 cgcatttaaa tccccacttc agaagttcct atactttcta gagaatagga acttcactat
     2521 agagtcgaat aagggcgaca ccccataatt agcccgggcg aaaggcccag tctttcgact
     2581 gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcatgggg agtccccaca
     2641 ctaccatcgg cgctacggcg tttcacttct gagttcggca tggggtcagg tgggaccacc
     2701 gcgctactgc cgccaggcaa acaaggggtg ttatgagcca tattcaggta taaatgggct
     2761 cgcgataatg ttcagaattg gttaattggt tgtaacactg acccctattt gtttattttt
     2821 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     2881 atattgaaaa aggaagaata tgagtattca acatttccgt gtcgccctta ttcccttttt
     2941 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     3001 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     3061 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     3121 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     3181 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     3241 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3301 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3361 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3421 cgagcgtgac accacgatgc ctgtagcgat ggcaacaacg ttgcgcaaac tattaactgg
     3481 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     3541 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatccgg
     3601 agccggtgag cgtggttctc gcggtatcat cgcagcgctg gggccagatg gtaagccctc
     3661 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     3721 gatcgctgag ataggtgcct cactgattaa gcattggtaa gcagagcatt acgctgactt
     3781 gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttacgcg cgcgtcgttc
     3841 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3901 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3961 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4021 aatactgttc ttctagtgta gccgtagtta gcccaccact tcaagaactc tgtagcaccg
     4081 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4141 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4201 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4261 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4321 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4381 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4441 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4501 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4561 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4621 cgcagcgagt cagtgagcga ggaagcggaa ggcgagagta gggaactgcc aggcatcaaa
     4681 ctaagcagaa ggcccctgac ggatggcctt tttgcgtttc tacaaactct ttctgtgttg
     4741 taaaacgacg gccagtctta agctcgggcc ccctgggcgg ttctgataac gagtaatcgt
     4801 taatccgcaa ataacgtaaa aacccgcttc ggcgggtttt tttatggggg gagtttaggg
     4861 aaagagcatt tgtcagaata tttaagggcg cctgtcactt tgcttgatat atgagaatta
     4921 tttaacctta taaatgagaa aaaagcaacg cactttaaat aagatacgtt gctttttcga
     4981 ttgatgaaca cctataatta aactattcat ctattattta tgattttttg tatatacaat
     5041 atttctagtt tgttaaagag aattaagaaa ataaatctcg aaaataataa agggaaaatc
     5101 agtttttgat atcaaaatta tacatgtcaa cgataataca aaatataata caaactataa
     5161 gatgttatca gtatttatta tgcatttaga ataaattttg tgtcgccctt attcgactca
     5221 ctatagaagt tcctattctc tagaaagtat aggaacttca cttcatttta tttaaatttg
     5281 cggccgcgct gaggtcttaa ttgagaatcc ggaccagcag ataatgcatg cagtggtgtt
     5341 ttgatgtata agcttgacat ttttttgctt ttcccgttat gagaaatgtt ttttttagac
     5401 tgaaaataaa agaatggata gtattaaaat agtaatgaac gtaataagga taggacagga
     5461 gcgataggaa aagatagaat tttggagctt tttttccctc tttttttgag caggagccaa
     5521 tagtagtcgg aaagatgagt gcatagattt ttcgtatttt atttcgagta aaaatatcac
     5581 gctcttaggt tcaataaaag aatatactat ttcttcaaaa tgcttctcta atctattttt
     5641 ttattttcct taataactga aatattattt tttgatgtaa aatgagtgaa caagaataag
     5701 ggatttttgg tatgtttgca agttctgtgg agttccatgt ataaggtata catattagaa
     5761 aagctttacg attatatact aacatcatgt acaaaacagt ttaataatga tctgtattgc
     5821 tggctcaatc cacgtaaatt aatgcctcag cactagtcct gcaggggtaa cgccagggtt
     5881 ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg actcactata
     5941 gggcgaattg ggtgcatagg ccactagtgg atctgatatc acctaataac ttcgtatagc
     6001 atacattata cgaagttata ttaagggttc tcgagagctc gttttattta ggttctatcg
     6061 aggagaaaaa gcgacaagaa gagatagacc atggataaat gattatgttc taaacactcc
     6121 tcagaagctc atcgaactgt catcctgcgt gaagattaaa atccaactta gaaatttcga
     6181 gcttacggag acaatcatat gggagaagca attggaagat agaaaaaagg tactcggtac
     6241 ataaatatat gtgattctgg gtagaagatc ggtctgcatt ggatggtggt aacgcatttt
     6301 tttacacaca ttacttgcct cgagcatcaa atggtggtta ttcgtggatc tatatcacgt
     6361 gatttgctta agaattgtcg ttcatggtga cacttttagc tttgacatga ttaagctcat
     6421 ctcaattgat gttatctaaa gtcatttcaa ctatctaaga tgtggttgtg attgggccat
     6481 tttgtgaaag ccagtacgcc agcgtcaata cactcccgtc aattagttgc accatgtcca
     6541 caaaatcata taccagtaga gctgagactc atgcaagtcc ggttgcatcg aaacttttac
     6601 gtttaatgga tgaaaagaag accaatttgt gtgcttctct tgacgttcgt tcgactgatg
     6661 agctattgaa acttgttgaa acgttgggtc catacatttg ccttttgaaa acacacgttg
     6721 atatcttgga tgatttcagt tatgagggta ctgtcgttcc attgaaagca ttggcagaga
     6781 aatacaagtt cttgatattt gaggacagaa aattcgccga tatcggtaac acagtcaaat
     6841 tacaatatac atcgggcgtt taccgtatcg cagaatggtc tgatatcacc aacgcccacg
     6901 gggttactgg tgctggtatt gttgctggct tgaaacaagg tgcgcaagag gtcaccaaag
     6961 aaccaagggg attattgatg cttgctgaat tgtcttccaa gggttctcta gcacacggtg
     7021 aatatactaa gggtaccgtt gatattgcaa agagtgataa agatttcgtt attgggttca
     7081 ttgctcagaa cgatatggga ggaagagaag aagggtttga ttggctaatc atgaccccag
     7141 gtgtaggttt agacgacaaa ggcgatgcat tgggtcagca gtacagaacc gtcgacgaag
     7201 ttgtaagtgg tggatcagat atcatcattg ttggcagagg acttttcgcc aagggtagag
     7261 atcctaaggt tgaaggtgaa agatacagaa atgctggatg ggaagcgt
//
