LOCUS       sequence-63465-e        6956 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6956)
  AUTHORS   Zordan RE, Ren Y, Pan SJ, Rotondo G, De Las Penas A, Iluore J, 
            Cormack BP
  TITLE     Expression plasmids for use in Candida glabrata.
  JOURNAL   G3 (Bethesda). 2013 Oct 3;3(10):1675-86. doi: 10.1534/g3.113.006908.
  PUBMED    23934995
REFERENCE   2  (bases 1 to 6956)
  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: "10.1534/g3.113"; 
            journalName: "G3 (Bethesda)"; date: "2013-10-3- 3"; volume: "3"; 
            issue: "10"; pages: "1675-86"
COMMENT     Sequence Label: pCU-PDC1-GFP
FEATURES             Location/Qualifiers
     source          1..6956
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        666..737
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(738..806,807..1598)
                     /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(956..975)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1769..2357
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2258..2277
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2511..2528
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2645..2666
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2681..2686,2687..2704,2705..2711)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2719..2735
                     /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)."
     primer_bind     2724..2746
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2743..2759
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2743..2759
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2778..2798
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2780..2798
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     3790..3806
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3790..3806
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3826..4542
                     /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"
     primer_bind     complement(4972..4991)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4973..4991)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5001..5018)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5001..5017)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5010..5032)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5158..5613
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5245..5264)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5455..5476
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5744..6547)
                     /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"
     promoter        complement(6548..6763)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6787..6806)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6906..6928
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagcaggt tagactaaca tgcaaaaagt ttatattcat ttttgtagtt attcttgttt
      121 tcattaggat gattacacac tgtggatgtg gattaatcga tcattgttaa tttagtgtag
      181 ctactaccat ttcaaacaaa acgtaaattc ttgcagtcgt actggatctg tgaatctatt
      241 agtatatatg aattaaagta gcttgtacat attattctgt tgaatcatat cgcagagcat
      301 tggttgaaat ccaaaatata aaaatgtaat atcacaaaaa ataatactaa ttctaacatt
      361 aatgggtcag atttttagtg aatacttaaa tttataatat cgtctattta agctagcaaa
      421 tggaacaaca tttaaagtaa gaacatcata tctacatgaa aatgtatatt tcaatctgac
      481 taataacgca gagcacatct ttcagtgatg tctgtcacat gatcaaaaag aattgtattt
      541 aaataattca taataaaagc ttaaaaaatt acaataatga aaataaagta aataatgaca
      601 tgggtaagag ttccgaaata gaatcttagt gtacaaaaca aaaattgcat cattagagat
      661 ccccacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
      721 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
      781 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
      841 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
      901 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
      961 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     1021 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     1081 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     1141 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     1201 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     1261 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     1321 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     1381 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     1441 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     1501 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     1561 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     1621 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     1681 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     1741 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     1801 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     1861 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     1921 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     1981 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2041 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2101 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     2161 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     2221 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     2281 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     2341 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     2401 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     2461 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     2521 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     2581 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     2641 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     2701 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     2761 atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg gagctcagca
     2821 tttttataca cgttttacgt attttttttg caatacacat agatacgtac gtacaaccct
     2881 ttctattgtg agaactttgt aacactcttt tgtagaccag tgtagtcagc aagcatcagc
     2941 aagcatcagc aagcatcagc aagcatcagc tagtctaaag cctttagcaa agtcaaaaac
     3001 caacacccac acaccgggat cccaccttcc ccgagatgcc tgttcccgac tcccagctgc
     3061 tcccacgacg gggaggaatc agaagacgga gatagaaaga gagcgttgaa cgcgcgcgat
     3121 gccaatgaga gataaaaaaa aagaaatacc aatcccacga catattctaa cgggctccac
     3181 agacaatgca caacaatcga gatgggattg aatcacagag agagagatca gaaaaaaaag
     3241 gaactgcaag ggtccccacc ccaaactccc ctccgctatc cactacaatc gccgcaccga
     3301 caaaatccat tgtgtccgca acgaccagag caaagagcca aaaacacaat tgaataacaa
     3361 agaaaagggc agagagccac ttagacaatt acgtaggaac catatttttg cataacttta
     3421 tgcaatttgc ccaaattcaa gtgaagcgag agaagccaga gagctacagc tgaagagcta
     3481 cagctgaagt gaaaagccaa gaatctctta gaatataccg tttggcttat ttgaatcaaa
     3541 tatttatgcg cttaaatccc ataaaaaagc aatttataca gataagtctg cagaaagaaa
     3601 aattttcttc ttgatacctg aacaaagaaa tcaaactcat caagaataaa tcaattcatg
     3661 aaaaaaaaac atatataaag gacaacatgg aatcaagttt caataatttt tagattgtac
     3721 ataaataaag agaccagact aatacaactg tataagctct aaacattcaa ttgccaaaaa
     3781 acattaacat ctagaactag tggatccccc gggctgcagg aattcatgtc taaaggtgaa
     3841 gaattattca ctggtgttgt cccaattttg gttgaattag atggtgatgt taatggtcac
     3901 aaattttctg tctccggtga aggtgaaggt gatgctactt acggtaaatt gaccttaaaa
     3961 tttatttgta ctactggtaa attgccggtt ccatggccaa ccttagtcac tactttcggt
     4021 tatggtgttc aatgttttgc tagataccca gatcatatga aacaacatga ctttttcaag
     4081 tctgccatgc cagaaggtta tgttcaagaa agaactattt ttttcaaaga tgacggtaac
     4141 tacaagacca gagctgaagt caagtttgaa ggtgatacct tagttaatag aatcgaatta
     4201 aaaggtattg attttaaaga agatggtaac attttaggtc acaaattgga atacaactat
     4261 aactctcaca atgtttacat catggctgac aaacaaaaga atggtatcaa agttaacttc
     4321 aaaattagac acaacattga agatggttct gttcaattag ctgaccatta tcaacaaaat
     4381 actccaattg gtgatggtcc agtcttgtta ccagacaacc attacttatc cactcaatct
     4441 gccttatcca aagatccaaa cgaaaagaga gaccacatgg tcttgttaga atttgttact
     4501 gctgctggta ttacccatgg tatggatgaa ttgtacaaat aactgcaggt cgacctcgag
     4561 aacacaaccc acagctacca ccatcaacaa tatttatata tataacgtac acatagaaat
     4621 cacacaaaca gagtatttat tcttaactac atgaactacc atcagaccgt ctgggccact
     4681 atataatgtg ccattcataa acgtgatcac tttacgtagc aggcaacccc aggtgaaaat
     4741 ttttcagcga gctgccagat tgtcaggtga aaaactgaaa aaaacttctg ggcgatgagc
     4801 ttgtggcggg aaattaagta tataaagcag ttagtttctt ctgcttcttg tggttctggg
     4861 tattcttgta atatttggaa gaatagaaaa tcaagactac tgcctttctt ttcatattat
     4921 agaggaagaa atacgcacga acacgatata gaggtaaagg tacccaattc gccctatagt
     4981 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     5041 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     5101 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggacg
     5161 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     5221 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     5281 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     5341 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     5401 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     5461 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     5521 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     5581 cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca
     5641 tctgtgcggt atttcacacc gcatagggta ataactgata taattaaatt gaagctctaa
     5701 tttgtgagtt tagtatacat gcatttactt ataatacagt tttttagttt tgctggccgc
     5761 atcttctcaa atatgcttcc cagcctgctt ttctgtaacg ttcaccctct accttagcat
     5821 cccttccctt tgcaaatagt cctcttccaa caataataat gtcagatcct gtagagacca
     5881 catcatccac ggttctatac tgttgaccca atgcgtctcc cttgtcatct aaacccacac
     5941 cgggtgtcat aatcaaccaa tcgtaacctt catctcttcc acccatgtct ctttgagcaa
     6001 taaagccgat aacaaaatct ttgtcgctct tcgcaatgtc aacagtaccc ttagtatatt
     6061 ctccagtaga tagggagccc ttgcatgaca attctgctaa catcaaaagg cctctaggtt
     6121 cctttgttac ttcttctgcc gcctgcttca aaccgctaac aatacctggg cccaccacac
     6181 cgtgtgcatt cgtaatgtct gcccattctg ctattctgta tacacccgca gagtactgca
     6241 atttgactgt attaccaatg tcagcaaatt ttctgtcttc gaagagtaaa aaattgtact
     6301 tggcggataa tgcctttagc ggcttaactg tgccctccat ggaaaaatca gtcaagatat
     6361 ccacatgtgt ttttagtaaa caaattttgg gacctaatgc ttcaactaac tccagtaatt
     6421 ccttggtggt acgaacatcc aatgaagcac acaagtttgt ttgcttttcg tgcatgatat
     6481 taaatagctt ggcagcaaca ggactaggat gagtagcagc acgttcctta tatgtagctt
     6541 tcgacatgat ttatcttcgt ttcctgcagg tttttgttct gtgcagttgg gttaagaata
     6601 ctgggcaatt tcatgtttct tcaacactac atatgcgtat atataccaat ctaagtctgt
     6661 gctccttcct tcgttcttcc ttctgttcgg agattaccga atcaaaaaaa tttcaaggaa
     6721 accgaaatca aaaaaaagaa taaaaaaaaa atgatgaatt gaaaaggtgg tatggtgcac
     6781 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     6841 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     6901 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcga
//
