LOCUS       sequence-63472-e        7276 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 7276)
  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 7276)
  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
FEATURES             Location/Qualifiers
     source          1..7276
                     /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"
     primer_bind     complement(4862..4878)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4863..4879)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(5292..5311)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5293..5311)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5321..5338)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5321..5337)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5330..5352)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5478..5933
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5565..5584)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5775..5796
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6064..6867)
                     /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(6868..7083)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7107..7126)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7226..7248
                     /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 ggaattcgat atcaagctta
     4861 tcgataccgt cgacctcgag aacacaaccc acagctacca ccatcaacaa tatttatata
     4921 tataacgtac acatagaaat cacacaaaca gagtatttat tcttaactac atgaactacc
     4981 atcagaccgt ctgggccact atataatgtg ccattcataa acgtgatcac tttacgtagc
     5041 aggcaacccc aggtgaaaat ttttcagcga gctgccagat tgtcaggtga aaaactgaaa
     5101 aaaacttctg ggcgatgagc ttgtggcggg aaattaagta tataaagcag ttagtttctt
     5161 ctgcttcttg tggttctggg tattcttgta atatttggaa gaatagaaaa tcaagactac
     5221 tgcctttctt ttcatattat agaggaagaa atacgcacga acacgatata gaggtaaagg
     5281 tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac
     5341 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     5401 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     5461 gcctgaatgg cgaatggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     5521 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     5581 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     5641 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     5701 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5761 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     5821 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     5881 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttcctgatg
     5941 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagggta ataactgata
     6001 taattaaatt gaagctctaa tttgtgagtt tagtatacat gcatttactt ataatacagt
     6061 tttttagttt tgctggccgc atcttctcaa atatgcttcc cagcctgctt ttctgtaacg
     6121 ttcaccctct accttagcat cccttccctt tgcaaatagt cctcttccaa caataataat
     6181 gtcagatcct gtagagacca catcatccac ggttctatac tgttgaccca atgcgtctcc
     6241 cttgtcatct aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt catctcttcc
     6301 acccatgtct ctttgagcaa taaagccgat aacaaaatct ttgtcgctct tcgcaatgtc
     6361 aacagtaccc ttagtatatt ctccagtaga tagggagccc ttgcatgaca attctgctaa
     6421 catcaaaagg cctctaggtt cctttgttac ttcttctgcc gcctgcttca aaccgctaac
     6481 aatacctggg cccaccacac cgtgtgcatt cgtaatgtct gcccattctg ctattctgta
     6541 tacacccgca gagtactgca atttgactgt attaccaatg tcagcaaatt ttctgtcttc
     6601 gaagagtaaa aaattgtact tggcggataa tgcctttagc ggcttaactg tgccctccat
     6661 ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa caaattttgg gacctaatgc
     6721 ttcaactaac tccagtaatt ccttggtggt acgaacatcc aatgaagcac acaagtttgt
     6781 ttgcttttcg tgcatgatat taaatagctt ggcagcaaca ggactaggat gagtagcagc
     6841 acgttcctta tatgtagctt tcgacatgat ttatcttcgt ttcctgcagg tttttgttct
     6901 gtgcagttgg gttaagaata ctgggcaatt tcatgtttct tcaacactac atatgcgtat
     6961 atataccaat ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg agattaccga
     7021 atcaaaaaaa tttcaaggaa accgaaatca aaaaaaagaa taaaaaaaaa atgatgaatt
     7081 gaaaaggtgg tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     7141 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     7201 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     7261 tcaccgaaac gcgcga
//
