LOCUS       sequence-63473-e        7978 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 7978)
  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 7978)
  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-ACO2-GFP
FEATURES             Location/Qualifiers
     source          1..7978
                     /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     4812..4828
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4812..4828
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             4848..5564
                     /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(5994..6013)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5995..6013)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6023..6040)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6023..6039)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6032..6054)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6180..6635
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6267..6286)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6477..6498
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6766..7569)
                     /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(7570..7785)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7809..7828)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7928..7950
                     /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 gagctcctgc
     2821 agtgtcccgt ttgtttcagt cctttgttat tatttcttga ataatggact ctcacttggg
     2881 aaaggaaata cttaagagtg gtattcaaac acctagttat atagtgattc cggttttgta
     2941 atcgtctaca atataagcaa agtttaggga atctagtttc attgttcttt tttttctttc
     3001 ttagtgttct ttattttgcc aatttttttt cttcttccca ttatgtggca ggaaggctca
     3061 attttcattt agcagagcta agaacgaaat tactgggggg acccgcttcg ttatctcgct
     3121 tttccctttt cctttttcct tctgcgtaac tttctaacag acttcttttc cctcgaggaa
     3181 agattttgtg cgtcgcttct ctccaaagtt gcttttttca ttttttgttc agttttcgca
     3241 gttccaagtg ctacgggaat cgtctctgcc aaatgtttgt gtgtgtgcta ctgcgcagat
     3301 cgttagggtg gaaaagaaag aaaatgccca ctgccttctc cgagaaaatc gtgtgaaaac
     3361 cgtgaaaacc gaaatggatc ttacctttta ggctcctgcg ttgcgctcta tgcatcggcc
     3421 tgccaattag aaatcccttc catacttttt ttctcttaaa tctgggaggg tggctacgcg
     3481 ggttcttctg catttgcttt tttctatcaa aaccaaatat caaatcttgc ggtgcttcct
     3541 gcagagaagt atctcgagga atagacattg tgagccaaca gaaaaaaaga gggggggagg
     3601 ccagagacga tggtggatgt gcgatgatct ctggcacctt gttaagaata gaactgtaag
     3661 tgctgtggga actgcgcaca gtcatggtgc tgggatatct aatttcgggt cggtggcaga
     3721 tcagacccac gtgggtctga tctaaagtaa agagcagaca ataacttgat agccagtgct
     3781 ttagttccag tcacactgtt aagcgggtac ccattgtatt tttacttgcc tggaggatgt
     3841 acggtaagtg tcaagcaaat attcttgagc aaaaaatttg catgaaaacg gatccatcct
     3901 ctgtgaattg acacataact ggagtagtaa ttagctttct atgtgcagac ggtgagacct
     3961 gacgcccatt agttagacaa tgtcgtatgg gccctctgca ctatgggggg gggactatgt
     4021 gactatttat ttgcgtacag tattacatgt atatatttct tgtgcatgtg gttcaagctg
     4081 tgttctttga aaaatgtaac ttaataagct tctatggcca agttggtaag gcgccacact
     4141 agtaatgtgg agatcatcag ttcaaatctg gttggaagca tttatttttt actttaaaaa
     4201 agcatgccat taataatcct actattggtg agaaaagcat tcgacttttt ggttttgata
     4261 ctatccaggt ttgtaacaca ggatactttc gtacacaccc ttttcgaata agttttttgc
     4321 ccaactacac aatcacccgt acaatgcagg ggatatccaa accattaagt agtcatcatt
     4381 ttgaaaaaat aaaagaaatc caaatttcca tcgggatttt tcttcagttt cctatcaacc
     4441 tctcatcttt gatctaacta gttctttaca aaaagctttc catttcatat cttcgggaaa
     4501 agggtttcct tttaggatgt gcaagaaaca tccattggct cttgactaat actaagaatt
     4561 ccagtgtaaa cagtaccacc ttcccattgt caattgaagt tggaaattaa gaaaaaattg
     4621 cggtgggaaa tatttggcga tgactaaaca tttattgcaa gtcctacaaa attttttttt
     4681 tcatagcaaa ctcactatat atattaacaa caataactta acaaataaaa ataacaaata
     4741 gaagtaaaac gtctatactg agccggacct tgtcttacat tacaacaaac tacaattcgt
     4801 acccactagc atctagaact agtggatccc ccgggctgca ggaattcatg tctaaaggtg
     4861 aagaattatt cactggtgtt gtcccaattt tggttgaatt agatggtgat gttaatggtc
     4921 acaaattttc tgtctccggt gaaggtgaag gtgatgctac ttacggtaaa ttgaccttaa
     4981 aatttatttg tactactggt aaattgccgg ttccatggcc aaccttagtc actactttcg
     5041 gttatggtgt tcaatgtttt gctagatacc cagatcatat gaaacaacat gactttttca
     5101 agtctgccat gccagaaggt tatgttcaag aaagaactat ttttttcaaa gatgacggta
     5161 actacaagac cagagctgaa gtcaagtttg aaggtgatac cttagttaat agaatcgaat
     5221 taaaaggtat tgattttaaa gaagatggta acattttagg tcacaaattg gaatacaact
     5281 ataactctca caatgtttac atcatggctg acaaacaaaa gaatggtatc aaagttaact
     5341 tcaaaattag acacaacatt gaagatggtt ctgttcaatt agctgaccat tatcaacaaa
     5401 atactccaat tggtgatggt ccagtcttgt taccagacaa ccattactta tccactcaat
     5461 ctgccttatc caaagatcca aacgaaaaga gagaccacat ggtcttgtta gaatttgtta
     5521 ctgctgctgg tattacccat ggtatggatg aattgtacaa ataactgcag gtcgacctcg
     5581 agaacacaac ccacagctac caccatcaac aatatttata tatataacgt acacatagaa
     5641 atcacacaaa cagagtattt attcttaact acatgaacta ccatcagacc gtctgggcca
     5701 ctatataatg tgccattcat aaacgtgatc actttacgta gcaggcaacc ccaggtgaaa
     5761 atttttcagc gagctgccag attgtcaggt gaaaaactga aaaaaacttc tgggcgatga
     5821 gcttgtggcg ggaaattaag tatataaagc agttagtttc ttctgcttct tgtggttctg
     5881 ggtattcttg taatatttgg aagaatagaa aatcaagact actgcctttc ttttcatatt
     5941 atagaggaag aaatacgcac gaacacgata tagaggtaaa ggtacccaat tcgccctata
     6001 gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     6061 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     6121 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga
     6181 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     6241 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     6301 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     6361 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     6421 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     6481 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     6541 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     6601 cgcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg
     6661 catctgtgcg gtatttcaca ccgcataggg taataactga tataattaaa ttgaagctct
     6721 aatttgtgag tttagtatac atgcatttac ttataataca gttttttagt tttgctggcc
     6781 gcatcttctc aaatatgctt cccagcctgc ttttctgtaa cgttcaccct ctaccttagc
     6841 atcccttccc tttgcaaata gtcctcttcc aacaataata atgtcagatc ctgtagagac
     6901 cacatcatcc acggttctat actgttgacc caatgcgtct cccttgtcat ctaaacccac
     6961 accgggtgtc ataatcaacc aatcgtaacc ttcatctctt ccacccatgt ctctttgagc
     7021 aataaagccg ataacaaaat ctttgtcgct cttcgcaatg tcaacagtac ccttagtata
     7081 ttctccagta gatagggagc ccttgcatga caattctgct aacatcaaaa ggcctctagg
     7141 ttcctttgtt acttcttctg ccgcctgctt caaaccgcta acaatacctg ggcccaccac
     7201 accgtgtgca ttcgtaatgt ctgcccattc tgctattctg tatacacccg cagagtactg
     7261 caatttgact gtattaccaa tgtcagcaaa ttttctgtct tcgaagagta aaaaattgta
     7321 cttggcggat aatgccttta gcggcttaac tgtgccctcc atggaaaaat cagtcaagat
     7381 atccacatgt gtttttagta aacaaatttt gggacctaat gcttcaacta actccagtaa
     7441 ttccttggtg gtacgaacat ccaatgaagc acacaagttt gtttgctttt cgtgcatgat
     7501 attaaatagc ttggcagcaa caggactagg atgagtagca gcacgttcct tatatgtagc
     7561 tttcgacatg atttatcttc gtttcctgca ggtttttgtt ctgtgcagtt gggttaagaa
     7621 tactgggcaa tttcatgttt cttcaacact acatatgcgt atatatacca atctaagtct
     7681 gtgctccttc cttcgttctt ccttctgttc ggagattacc gaatcaaaaa aatttcaagg
     7741 aaaccgaaat caaaaaaaag aataaaaaaa aaatgatgaa ttgaaaaggt ggtatggtgc
     7801 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     7861 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     7921 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcga
//
