LOCUS       sequence-63487-e        6725 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 6725)
  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 6725)
  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-MET3-GFP
FEATURES             Location/Qualifiers
     source          1..6725
                     /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     2680..2696
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2680..2696
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             2716..3432
                     /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(3906..3922)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3906..3922)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3919..3941)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3930..3946
                     /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(3954..3960,3961..3978,3979..3984))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3999..4020
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4137..4154)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4308..4896)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4388..4407)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5067..5858,5859..5927))
                     /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     5690..5709
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5928..6032)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6698..6716
                     /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 ctcataccag ttacaattag tattacaatg
     1681 gtttaccaag tacaaattat gtaatatggt tataattggc ttatatgtat ggaggtaggg
     1741 aactgcaggt tattgttctg ttaaactacc agtaatgcaa gacggctaag tcatatgact
     1801 gccactttgt caatcctgaa gaaaatgaca atagaaagtg atataataca cgtgatatgc
     1861 gtaggatggg acccaaagat agccacatca catgatcttc aaaaaccccg tcaaaaagga
     1921 ataaatcgaa ggaaaaaaaa tgccacgtga ctttgatagt caaaatcagg ttatattact
     1981 atacgatccc ccacacataa cctttaccac acaccgcacg ggctggactt atcctattag
     2041 aaagcggtgc agccagggcc aagaaaacgc gaacgacgca gaaaaaacgt caagcaaaaa
     2101 aactgtggtg ttatttaaag catacaattt cctgcctccc tttaatgcta cgggggatct
     2161 agcaaatggg aaaatatcac atgactttgg ctagaagggg tgggaaaaca gatattttag
     2221 tcacattgtt tgatttcacg tactacacga tacactacac tacacaatac actgcaatac
     2281 actacactgt atggttccct ccggccctgg atatgctagc gaaggatccc aagcccatcg
     2341 aggaaatcat tcaagagcat ctgcaggtct caaatacact aagtccaaac acaaacagca
     2401 ctgagctaat ccaagacacc aattgcatgc cttctcaatt agaacagttt attacacagc
     2461 ttataactgt gcatcttgcc attcttttcg agatagccat ttgcatttga ttaacatgct
     2521 tagctcgttc tcaagccaca gtaaaatgga ttgccttttg agtttccatc gttgtataaa
     2581 taggcactca ttcctatagc cttgctcttt ggtcttgtct ctgtaatggt aatagctgta
     2641 agtcaggata taacactcca gaaaagaaac acctaacaat ctagaactag tggatccccc
     2701 gggctgcagg aattcatgtc taaaggtgaa gaattattca ctggtgttgt cccaattttg
     2761 gttgaattag atggtgatgt taatggtcac aaattttctg tctccggtga aggtgaaggt
     2821 gatgctactt acggtaaatt gaccttaaaa tttatttgta ctactggtaa attgccggtt
     2881 ccatggccaa ccttagtcac tactttcggt tatggtgttc aatgttttgc tagataccca
     2941 gatcatatga aacaacatga ctttttcaag tctgccatgc cagaaggtta tgttcaagaa
     3001 agaactattt ttttcaaaga tgacggtaac tacaagacca gagctgaagt caagtttgaa
     3061 ggtgatacct tagttaatag aatcgaatta aaaggtattg attttaaaga agatggtaac
     3121 attttaggtc acaaattgga atacaactat aactctcaca atgtttacat catggctgac
     3181 aaacaaaaga atggtatcaa agttaacttc aaaattagac acaacattga agatggttct
     3241 gttcaattag ctgaccatta tcaacaaaat actccaattg gtgatggtcc agtcttgtta
     3301 ccagacaacc attacttatc cactcaatct gccttatcca aagatccaaa cgaaaagaga
     3361 gaccacatgg tcttgttaga atttgttact gctgctggta ttacccatgg tatggatgaa
     3421 ttgtacaaat aactgcaggt cgacctcgag aacacaaccc acagctacca ccatcaacaa
     3481 tatttatata tataacgtac acatagaaat cacacaaaca gagtatttat tcttaactac
     3541 atgaactacc atcagaccgt ctgggccact atataatgtg ccattcataa acgtgatcac
     3601 tttacgtagc aggcaacccc aggtgaaaat ttttcagcga gctgccagat tgtcaggtga
     3661 aaaactgaaa aaaacttctg ggcgatgagc ttgtggcggg aaattaagta tataaagcag
     3721 ttagtttctt ctgcttcttg tggttctggg tattcttgta atatttggaa gaatagaaaa
     3781 tcaagactac tgcctttctt ttcatattat agaggaagaa atacgcacga acacgatata
     3841 gaggtaaagg tacccgggga tcctctagag tcgacctgca ggcatgcaag cttggcgtaa
     3901 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     3961 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4021 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4081 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4141 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4201 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4261 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4321 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4381 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4441 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4501 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4561 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4621 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4681 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4741 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4801 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4861 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4921 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4981 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5041 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5101 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5161 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5221 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5281 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5341 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5401 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5461 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5521 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5581 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5641 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5701 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5761 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5821 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5881 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5941 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6001 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     6061 gtcgggatct ctaatgatgc aatttttgtt ttgtacacta agattctatt tcggaactct
     6121 tacccatgtc attatttact ttattttcat tattgtaatt ttttaagctt ttattatgaa
     6181 ttatttaaat acaattcttt ttgatcatgt gacagacatc actgaaagat gtgctctgcg
     6241 ttattagtca gattgaaata tacattttca tgtagatatg atgttcttac tttaaatgtt
     6301 gttccatttg ctagcttaaa tagacgatat tataaattta agtattcact aaaaatctga
     6361 cccattaatg ttagaattag tattattttt tgtgatatta catttttata ttttggattt
     6421 caaccaatgc tctgcgatat gattcaacag aataatatgt acaagctact ttaattcata
     6481 tatactaata gattcacaga tccagtacga ctgcaagaat ttacgttttg tttgaaatgg
     6541 tagtagctac actaaattaa caatgatcga ttaatccaca tccacagtgt gtaatcatcc
     6601 taatgaaaac aagaataact acaaaaatga atataaactt tttgcatgtt agtctgacgt
     6661 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     6721 tcgtc
//
