LOCUS       sequence-63470-e        6672 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 6672)
  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 6672)
  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-PDC1-GFP
FEATURES             Location/Qualifiers
     source          1..6672
                     /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     2627..2643
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2627..2643
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             2663..3379
                     /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(3853..3869)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3853..3869)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3866..3888)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3877..3893
                     /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(3901..3907,3908..3925,3926..3931))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3946..3967
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4084..4101)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4255..4843)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4335..4354)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5014..5805,5806..5874))
                     /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     5637..5656
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5875..5979)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6645..6663
                     /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 ctcagcattt ttatacacgt tttacgtatt
     1681 ttttttgcaa tacacataga tacgtacgta caaccctttc tattgtgaga actttgtaac
     1741 actcttttgt agaccagtgt agtcagcaag catcagcaag catcagcaag catcagcaag
     1801 catcagctag tctaaagcct ttagcaaagt caaaaaccaa cacccacaca ccgggatccc
     1861 accttccccg agatgcctgt tcccgactcc cagctgctcc cacgacgggg aggaatcaga
     1921 agacggagat agaaagagag cgttgaacgc gcgcgatgcc aatgagagat aaaaaaaaag
     1981 aaataccaat cccacgacat attctaacgg gctccacaga caatgcacaa caatcgagat
     2041 gggattgaat cacagagaga gagatcagaa aaaaaaggaa ctgcaagggt ccccacccca
     2101 aactcccctc cgctatccac tacaatcgcc gcaccgacaa aatccattgt gtccgcaacg
     2161 accagagcaa agagccaaaa acacaattga ataacaaaga aaagggcaga gagccactta
     2221 gacaattacg taggaaccat atttttgcat aactttatgc aatttgccca aattcaagtg
     2281 aagcgagaga agccagagag ctacagctga agagctacag ctgaagtgaa aagccaagaa
     2341 tctcttagaa tataccgttt ggcttatttg aatcaaatat ttatgcgctt aaatcccata
     2401 aaaaagcaat ttatacagat aagtctgcag aaagaaaaat tttcttcttg atacctgaac
     2461 aaagaaatca aactcatcaa gaataaatca attcatgaaa aaaaaacata tataaaggac
     2521 aacatggaat caagtttcaa taatttttag attgtacata aataaagaga ccagactaat
     2581 acaactgtat aagctctaaa cattcaattg ccaaaaaaca ttaacatcta gaactagtgg
     2641 atcccccggg ctgcaggaat tcatgtctaa aggtgaagaa ttattcactg gtgttgtccc
     2701 aattttggtt gaattagatg gtgatgttaa tggtcacaaa ttttctgtct ccggtgaagg
     2761 tgaaggtgat gctacttacg gtaaattgac cttaaaattt atttgtacta ctggtaaatt
     2821 gccggttcca tggccaacct tagtcactac tttcggttat ggtgttcaat gttttgctag
     2881 atacccagat catatgaaac aacatgactt tttcaagtct gccatgccag aaggttatgt
     2941 tcaagaaaga actatttttt tcaaagatga cggtaactac aagaccagag ctgaagtcaa
     3001 gtttgaaggt gataccttag ttaatagaat cgaattaaaa ggtattgatt ttaaagaaga
     3061 tggtaacatt ttaggtcaca aattggaata caactataac tctcacaatg tttacatcat
     3121 ggctgacaaa caaaagaatg gtatcaaagt taacttcaaa attagacaca acattgaaga
     3181 tggttctgtt caattagctg accattatca acaaaatact ccaattggtg atggtccagt
     3241 cttgttacca gacaaccatt acttatccac tcaatctgcc ttatccaaag atccaaacga
     3301 aaagagagac cacatggtct tgttagaatt tgttactgct gctggtatta cccatggtat
     3361 ggatgaattg tacaaataac tgcaggtcga cctcgagaac acaacccaca gctaccacca
     3421 tcaacaatat ttatatatat aacgtacaca tagaaatcac acaaacagag tatttattct
     3481 taactacatg aactaccatc agaccgtctg ggccactata taatgtgcca ttcataaacg
     3541 tgatcacttt acgtagcagg caaccccagg tgaaaatttt tcagcgagct gccagattgt
     3601 caggtgaaaa actgaaaaaa acttctgggc gatgagcttg tggcgggaaa ttaagtatat
     3661 aaagcagtta gtttcttctg cttcttgtgg ttctgggtat tcttgtaata tttggaagaa
     3721 tagaaaatca agactactgc ctttcttttc atattataga ggaagaaata cgcacgaaca
     3781 cgatatagag gtaaaggtac ccggggatcc tctagagtcg acctgcaggc atgcaagctt
     3841 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     3901 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     3961 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4021 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4081 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4141 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4201 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4261 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4321 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4381 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4441 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4501 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4561 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4621 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4681 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4741 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4801 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4861 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4921 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4981 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5041 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5101 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5161 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5221 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5281 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5341 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5401 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5461 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5521 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5581 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5641 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5701 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5761 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5821 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5881 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5941 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6001 acctgacgtc gggatctcta atgatgcaat ttttgttttg tacactaaga ttctatttcg
     6061 gaactcttac ccatgtcatt atttacttta ttttcattat tgtaattttt taagctttta
     6121 ttatgaatta tttaaataca attctttttg atcatgtgac agacatcact gaaagatgtg
     6181 ctctgcgtta ttagtcagat tgaaatatac attttcatgt agatatgatg ttcttacttt
     6241 aaatgttgtt ccatttgcta gcttaaatag acgatattat aaatttaagt attcactaaa
     6301 aatctgaccc attaatgtta gaattagtat tattttttgt gatattacat ttttatattt
     6361 tggatttcaa ccaatgctct gcgatatgat tcaacagaat aatatgtaca agctacttta
     6421 attcatatat actaatagat tcacagatcc agtacgactg caagaattta cgttttgttt
     6481 gaaatggtag tagctacact aaattaacaa tgatcgatta atccacatcc acagtgtgta
     6541 atcatcctaa tgaaaacaag aataactaca aaaatgaata taaacttttt gcatgttagt
     6601 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     6661 ggccctttcg tc
//
