LOCUS       sequence-63484-e        7009 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 7009)
  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 7009)
  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-MET3-GFP
FEATURES             Location/Qualifiers
     source          1..7009
                     /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     3843..3859
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3843..3859
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3879..4595
                     /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(5025..5044)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5026..5044)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5054..5071)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5054..5070)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5063..5085)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5211..5666
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5298..5317)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5508..5529
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5797..6600)
                     /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(6601..6816)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6840..6859)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6959..6981
                     /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 gagctcatac
     2821 cagttacaat tagtattaca atggtttacc aagtacaaat tatgtaatat ggttataatt
     2881 ggcttatatg tatggaggta gggaactgca ggttattgtt ctgttaaact accagtaatg
     2941 caagacggct aagtcatatg actgccactt tgtcaatcct gaagaaaatg acaatagaaa
     3001 gtgatataat acacgtgata tgcgtaggat gggacccaaa gatagccaca tcacatgatc
     3061 ttcaaaaacc ccgtcaaaaa ggaataaatc gaaggaaaaa aaatgccacg tgactttgat
     3121 agtcaaaatc aggttatatt actatacgat cccccacaca taacctttac cacacaccgc
     3181 acgggctgga cttatcctat tagaaagcgg tgcagccagg gccaagaaaa cgcgaacgac
     3241 gcagaaaaaa cgtcaagcaa aaaaactgtg gtgttattta aagcatacaa tttcctgcct
     3301 ccctttaatg ctacggggga tctagcaaat gggaaaatat cacatgactt tggctagaag
     3361 gggtgggaaa acagatattt tagtcacatt gtttgatttc acgtactaca cgatacacta
     3421 cactacacaa tacactgcaa tacactacac tgtatggttc cctccggccc tggatatgct
     3481 agcgaaggat cccaagccca tcgaggaaat cattcaagag catctgcagg tctcaaatac
     3541 actaagtcca aacacaaaca gcactgagct aatccaagac accaattgca tgccttctca
     3601 attagaacag tttattacac agcttataac tgtgcatctt gccattcttt tcgagatagc
     3661 catttgcatt tgattaacat gcttagctcg ttctcaagcc acagtaaaat ggattgcctt
     3721 ttgagtttcc atcgttgtat aaataggcac tcattcctat agccttgctc tttggtcttg
     3781 tctctgtaat ggtaatagct gtaagtcagg atataacact ccagaaaaga aacacctaac
     3841 aatctagaac tagtggatcc cccgggctgc aggaattcat gtctaaaggt gaagaattat
     3901 tcactggtgt tgtcccaatt ttggttgaat tagatggtga tgttaatggt cacaaatttt
     3961 ctgtctccgg tgaaggtgaa ggtgatgcta cttacggtaa attgacctta aaatttattt
     4021 gtactactgg taaattgccg gttccatggc caaccttagt cactactttc ggttatggtg
     4081 ttcaatgttt tgctagatac ccagatcata tgaaacaaca tgactttttc aagtctgcca
     4141 tgccagaagg ttatgttcaa gaaagaacta tttttttcaa agatgacggt aactacaaga
     4201 ccagagctga agtcaagttt gaaggtgata ccttagttaa tagaatcgaa ttaaaaggta
     4261 ttgattttaa agaagatggt aacattttag gtcacaaatt ggaatacaac tataactctc
     4321 acaatgttta catcatggct gacaaacaaa agaatggtat caaagttaac ttcaaaatta
     4381 gacacaacat tgaagatggt tctgttcaat tagctgacca ttatcaacaa aatactccaa
     4441 ttggtgatgg tccagtcttg ttaccagaca accattactt atccactcaa tctgccttat
     4501 ccaaagatcc aaacgaaaag agagaccaca tggtcttgtt agaatttgtt actgctgctg
     4561 gtattaccca tggtatggat gaattgtaca aataactgca ggtcgacctc gagaacacaa
     4621 cccacagcta ccaccatcaa caatatttat atatataacg tacacataga aatcacacaa
     4681 acagagtatt tattcttaac tacatgaact accatcagac cgtctgggcc actatataat
     4741 gtgccattca taaacgtgat cactttacgt agcaggcaac cccaggtgaa aatttttcag
     4801 cgagctgcca gattgtcagg tgaaaaactg aaaaaaactt ctgggcgatg agcttgtggc
     4861 gggaaattaa gtatataaag cagttagttt cttctgcttc ttgtggttct gggtattctt
     4921 gtaatatttg gaagaataga aaatcaagac tactgccttt cttttcatat tatagaggaa
     4981 gaaatacgca cgaacacgat atagaggtaa aggtacccaa ttcgccctat agtgagtcgt
     5041 attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     5101 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     5161 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg acgcgccctg
     5221 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     5281 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     5341 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     5401 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     5461 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     5521 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     5581 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     5641 taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc
     5701 ggtatttcac accgcatagg gtaataactg atataattaa attgaagctc taatttgtga
     5761 gtttagtata catgcattta cttataatac agttttttag ttttgctggc cgcatcttct
     5821 caaatatgct tcccagcctg cttttctgta acgttcaccc tctaccttag catcccttcc
     5881 ctttgcaaat agtcctcttc caacaataat aatgtcagat cctgtagaga ccacatcatc
     5941 cacggttcta tactgttgac ccaatgcgtc tcccttgtca tctaaaccca caccgggtgt
     6001 cataatcaac caatcgtaac cttcatctct tccacccatg tctctttgag caataaagcc
     6061 gataacaaaa tctttgtcgc tcttcgcaat gtcaacagta cccttagtat attctccagt
     6121 agatagggag cccttgcatg acaattctgc taacatcaaa aggcctctag gttcctttgt
     6181 tacttcttct gccgcctgct tcaaaccgct aacaatacct gggcccacca caccgtgtgc
     6241 attcgtaatg tctgcccatt ctgctattct gtatacaccc gcagagtact gcaatttgac
     6301 tgtattacca atgtcagcaa attttctgtc ttcgaagagt aaaaaattgt acttggcgga
     6361 taatgccttt agcggcttaa ctgtgccctc catggaaaaa tcagtcaaga tatccacatg
     6421 tgtttttagt aaacaaattt tgggacctaa tgcttcaact aactccagta attccttggt
     6481 ggtacgaaca tccaatgaag cacacaagtt tgtttgcttt tcgtgcatga tattaaatag
     6541 cttggcagca acaggactag gatgagtagc agcacgttcc ttatatgtag ctttcgacat
     6601 gatttatctt cgtttcctgc aggtttttgt tctgtgcagt tgggttaaga atactgggca
     6661 atttcatgtt tcttcaacac tacatatgcg tatatatacc aatctaagtc tgtgctcctt
     6721 ccttcgttct tccttctgtt cggagattac cgaatcaaaa aaatttcaag gaaaccgaaa
     6781 tcaaaaaaaa gaataaaaaa aaaatgatga attgaaaagg tggtatggtg cactctcagt
     6841 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     6901 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     6961 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcga
//
