LOCUS       sequence-63476-e        7694 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 7694)
  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 7694)
  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: pCN-ACO2-GFP
FEATURES             Location/Qualifiers
     source          1..7694
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        180..585
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             599..1171
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARELGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      1178..1425
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1194..1212)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     1610..1632
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1624..1641
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1625..1641
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3649..3665
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3649..3665
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3685..4401
                     /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(4875..4891)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4875..4891)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4888..4910)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4899..4915
                     /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)."
     promoter        complement(join(4923..4929,4930..4947,4948..4953))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4968..4989
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5106..5123)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5277..5865)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5357..5376)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6036..6827,6828..6896))
                     /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     6659..6678
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6897..7001)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7667..7685
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccat agcttcaaaa tgtttctact ccttttttac tcttccagat tttctcggac
      241 tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat ttcccctctt
      301 tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa aaaagagacc
      361 gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg tttctttttc
      421 ttgaaaattt ttttttgatt tttttctctt tcgatgacct cccattgata tttaagttaa
      481 taaacggtct tcaatttctc aagtttcagt ttcatttttc ttgttctatt acaacttttt
      541 ttacttcttg ctcattagaa agaaagcata gcaatctaat ctaagttttg gatctaaaat
      601 gggcaccact cttgacgaca cggcttaccg ataccgcacc agtgtcccgg gggacgccga
      661 ggccatcgag gcactggatg ggtccttcac caccgacacc gtcttccgcg tcaccgccac
      721 cggggacggc ttcaccctgc gggaggtgcc ggtggacccg cccctgacca aggtgttccc
      781 cgacgacgaa tcggacgacg aatcggacga cggggaggac ggcgaccctg actcccggac
      841 gttcgtcgcg tacggggacg acggcgacct ggcgggcttc gtggtcgtct cgtactccgg
      901 ctggaaccgc cggctgaccg tcgaggacat cgaggtcgcc ccggagcacc gggggcacgg
      961 ggtcgggcgc gcgttgatgg ggctcgcgac ggagttcgcc cgcgagctgg gcgccgggca
     1021 cctctggctg gaggtcacca acgtcaacgc accggcgatc cacgcgtacc ggcggatggg
     1081 gttcaccctc tgcggcctgg acaccgccct gtacgacggc accgcctcgg acggcgagca
     1141 ggcgctctac atgagcatgc cctgccccta agtcgagtca tgtaattagt tatgtcacgc
     1201 ttacattcac gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct
     1261 gaagtctagg tccctattta tttttttata gttatgttag tattaagaac gttatttata
     1321 tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa
     1381 accttgcttg agaaggtttt gggacgctcg aaggctttaa tttgcggccg tatgcggtgt
     1441 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg
     1501 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     1561 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     1621 cgttgtaaaa cgacggccag tgaattcgag ctcctgcagt gtcccgtttg tttcagtcct
     1681 ttgttattat ttcttgaata atggactctc acttgggaaa ggaaatactt aagagtggta
     1741 ttcaaacacc tagttatata gtgattccgg ttttgtaatc gtctacaata taagcaaagt
     1801 ttagggaatc tagtttcatt gttctttttt ttctttctta gtgttcttta ttttgccaat
     1861 tttttttctt cttcccatta tgtggcagga aggctcaatt ttcatttagc agagctaaga
     1921 acgaaattac tggggggacc cgcttcgtta tctcgctttt cccttttcct ttttccttct
     1981 gcgtaacttt ctaacagact tcttttccct cgaggaaaga ttttgtgcgt cgcttctctc
     2041 caaagttgct tttttcattt tttgttcagt tttcgcagtt ccaagtgcta cgggaatcgt
     2101 ctctgccaaa tgtttgtgtg tgtgctactg cgcagatcgt tagggtggaa aagaaagaaa
     2161 atgcccactg ccttctccga gaaaatcgtg tgaaaaccgt gaaaaccgaa atggatctta
     2221 ccttttaggc tcctgcgttg cgctctatgc atcggcctgc caattagaaa tcccttccat
     2281 actttttttc tcttaaatct gggagggtgg ctacgcgggt tcttctgcat ttgctttttt
     2341 ctatcaaaac caaatatcaa atcttgcggt gcttcctgca gagaagtatc tcgaggaata
     2401 gacattgtga gccaacagaa aaaaagaggg ggggaggcca gagacgatgg tggatgtgcg
     2461 atgatctctg gcaccttgtt aagaatagaa ctgtaagtgc tgtgggaact gcgcacagtc
     2521 atggtgctgg gatatctaat ttcgggtcgg tggcagatca gacccacgtg ggtctgatct
     2581 aaagtaaaga gcagacaata acttgatagc cagtgcttta gttccagtca cactgttaag
     2641 cgggtaccca ttgtattttt acttgcctgg aggatgtacg gtaagtgtca agcaaatatt
     2701 cttgagcaaa aaatttgcat gaaaacggat ccatcctctg tgaattgaca cataactgga
     2761 gtagtaatta gctttctatg tgcagacggt gagacctgac gcccattagt tagacaatgt
     2821 cgtatgggcc ctctgcacta tggggggggg actatgtgac tatttatttg cgtacagtat
     2881 tacatgtata tatttcttgt gcatgtggtt caagctgtgt tctttgaaaa atgtaactta
     2941 ataagcttct atggccaagt tggtaaggcg ccacactagt aatgtggaga tcatcagttc
     3001 aaatctggtt ggaagcattt attttttact ttaaaaaagc atgccattaa taatcctact
     3061 attggtgaga aaagcattcg actttttggt tttgatacta tccaggtttg taacacagga
     3121 tactttcgta cacacccttt tcgaataagt tttttgccca actacacaat cacccgtaca
     3181 atgcagggga tatccaaacc attaagtagt catcattttg aaaaaataaa agaaatccaa
     3241 atttccatcg ggatttttct tcagtttcct atcaacctct catctttgat ctaactagtt
     3301 ctttacaaaa agctttccat ttcatatctt cgggaaaagg gtttcctttt aggatgtgca
     3361 agaaacatcc attggctctt gactaatact aagaattcca gtgtaaacag taccaccttc
     3421 ccattgtcaa ttgaagttgg aaattaagaa aaaattgcgg tgggaaatat ttggcgatga
     3481 ctaaacattt attgcaagtc ctacaaaatt ttttttttca tagcaaactc actatatata
     3541 ttaacaacaa taacttaaca aataaaaata acaaatagaa gtaaaacgtc tatactgagc
     3601 cggaccttgt cttacattac aacaaactac aattcgtacc cactagcatc tagaactagt
     3661 ggatcccccg ggctgcagga attcatgtct aaaggtgaag aattattcac tggtgttgtc
     3721 ccaattttgg ttgaattaga tggtgatgtt aatggtcaca aattttctgt ctccggtgaa
     3781 ggtgaaggtg atgctactta cggtaaattg accttaaaat ttatttgtac tactggtaaa
     3841 ttgccggttc catggccaac cttagtcact actttcggtt atggtgttca atgttttgct
     3901 agatacccag atcatatgaa acaacatgac tttttcaagt ctgccatgcc agaaggttat
     3961 gttcaagaaa gaactatttt tttcaaagat gacggtaact acaagaccag agctgaagtc
     4021 aagtttgaag gtgatacctt agttaataga atcgaattaa aaggtattga ttttaaagaa
     4081 gatggtaaca ttttaggtca caaattggaa tacaactata actctcacaa tgtttacatc
     4141 atggctgaca aacaaaagaa tggtatcaaa gttaacttca aaattagaca caacattgaa
     4201 gatggttctg ttcaattagc tgaccattat caacaaaata ctccaattgg tgatggtcca
     4261 gtcttgttac cagacaacca ttacttatcc actcaatctg ccttatccaa agatccaaac
     4321 gaaaagagag accacatggt cttgttagaa tttgttactg ctgctggtat tacccatggt
     4381 atggatgaat tgtacaaata actgcaggtc gacctcgaga acacaaccca cagctaccac
     4441 catcaacaat atttatatat ataacgtaca catagaaatc acacaaacag agtatttatt
     4501 cttaactaca tgaactacca tcagaccgtc tgggccacta tataatgtgc cattcataaa
     4561 cgtgatcact ttacgtagca ggcaacccca ggtgaaaatt tttcagcgag ctgccagatt
     4621 gtcaggtgaa aaactgaaaa aaacttctgg gcgatgagct tgtggcggga aattaagtat
     4681 ataaagcagt tagtttcttc tgcttcttgt ggttctgggt attcttgtaa tatttggaag
     4741 aatagaaaat caagactact gcctttcttt tcatattata gaggaagaaa tacgcacgaa
     4801 cacgatatag aggtaaaggt acccggggat cctctagagt cgacctgcag gcatgcaagc
     4861 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     4921 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     4981 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5041 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5101 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5161 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5221 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5281 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5341 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5401 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5461 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5521 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5581 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5641 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5701 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5761 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5821 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5881 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5941 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6001 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6061 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6121 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6181 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6241 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6301 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6361 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6421 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6481 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6541 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6601 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6661 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6721 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6781 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6841 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6901 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6961 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7021 ccacctgacg tcgggatctc taatgatgca atttttgttt tgtacactaa gattctattt
     7081 cggaactctt acccatgtca ttatttactt tattttcatt attgtaattt tttaagcttt
     7141 tattatgaat tatttaaata caattctttt tgatcatgtg acagacatca ctgaaagatg
     7201 tgctctgcgt tattagtcag attgaaatat acattttcat gtagatatga tgttcttact
     7261 ttaaatgttg ttccatttgc tagcttaaat agacgatatt ataaatttaa gtattcacta
     7321 aaaatctgac ccattaatgt tagaattagt attatttttt gtgatattac atttttatat
     7381 tttggatttc aaccaatgct ctgcgatatg attcaacaga ataatatgta caagctactt
     7441 taattcatat atactaatag attcacagat ccagtacgac tgcaagaatt tacgttttgt
     7501 ttgaaatggt agtagctaca ctaaattaac aatgatcgat taatccacat ccacagtgtg
     7561 taatcatcct aatgaaaaca agaataacta caaaaatgaa tataaacttt ttgcatgtta
     7621 gtctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     7681 gaggcccttt cgtc
//
