LOCUS       sequence-63415-e        6043 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 6043)
  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 6043)
  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-EGD2-GFP
FEATURES             Location/Qualifiers
     source          1..6043
                     /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     1998..2014
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1998..2014
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             2034..2750
                     /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(3224..3240)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3224..3240)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3237..3259)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3248..3264
                     /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(3272..3278,3279..3296,3297..3302))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3317..3338
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3455..3472)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3626..4214)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3706..3725)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4385..5176,5177..5245))
                     /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     5008..5027
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5246..5350)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6016..6034
                     /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 ctctgtccac ttcactcacc agtaataagt
     1681 tgcccagtca ccatccctta tatacgctgt ggctaccgag atggccgaac ccactccaaa
     1741 tatatgggat actcaaactg aaagatgatg atgagcttat tatggtaatg ctgtacgtac
     1801 attacccccg tggcaagaaa cacgtacaac gctacatttc tcgtcgacgc tttttttttt
     1861 caaaatttct ttcatcgaag agctttggag ggtctcatag cacatataag ttgttattgc
     1921 tacacaattt tggtcaattg agtagctgtt gcaataggga caaatcaaaa acacgtcaat
     1981 aatacagata tacaaagtct agaactagtg gatcccccgg gctgcaggaa ttcatgtcta
     2041 aaggtgaaga attattcact ggtgttgtcc caattttggt tgaattagat ggtgatgtta
     2101 atggtcacaa attttctgtc tccggtgaag gtgaaggtga tgctacttac ggtaaattga
     2161 ccttaaaatt tatttgtact actggtaaat tgccggttcc atggccaacc ttagtcacta
     2221 ctttcggtta tggtgttcaa tgttttgcta gatacccaga tcatatgaaa caacatgact
     2281 ttttcaagtc tgccatgcca gaaggttatg ttcaagaaag aactattttt ttcaaagatg
     2341 acggtaacta caagaccaga gctgaagtca agtttgaagg tgatacctta gttaatagaa
     2401 tcgaattaaa aggtattgat tttaaagaag atggtaacat tttaggtcac aaattggaat
     2461 acaactataa ctctcacaat gtttacatca tggctgacaa acaaaagaat ggtatcaaag
     2521 ttaacttcaa aattagacac aacattgaag atggttctgt tcaattagct gaccattatc
     2581 aacaaaatac tccaattggt gatggtccag tcttgttacc agacaaccat tacttatcca
     2641 ctcaatctgc cttatccaaa gatccaaacg aaaagagaga ccacatggtc ttgttagaat
     2701 ttgttactgc tgctggtatt acccatggta tggatgaatt gtacaaataa ctgcaggtcg
     2761 acctcgagaa cacaacccac agctaccacc atcaacaata tttatatata taacgtacac
     2821 atagaaatca cacaaacaga gtatttattc ttaactacat gaactaccat cagaccgtct
     2881 gggccactat ataatgtgcc attcataaac gtgatcactt tacgtagcag gcaaccccag
     2941 gtgaaaattt ttcagcgagc tgccagattg tcaggtgaaa aactgaaaaa aacttctggg
     3001 cgatgagctt gtggcgggaa attaagtata taaagcagtt agtttcttct gcttcttgtg
     3061 gttctgggta ttcttgtaat atttggaaga atagaaaatc aagactactg cctttctttt
     3121 catattatag aggaagaaat acgcacgaac acgatataga ggtaaaggta cccggggatc
     3181 ctctagagtc gacctgcagg catgcaagct tggcgtaatc atggtcatag ctgtttcctg
     3241 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3301 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3361 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3421 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3481 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3541 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3601 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3661 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3721 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3781 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3841 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3901 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3961 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4021 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4081 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4141 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4201 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4261 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4321 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4381 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4441 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4501 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4561 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4621 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4681 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4741 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4801 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4861 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4921 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4981 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5041 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5101 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5161 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5221 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5281 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5341 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgggatctct aatgatgcaa
     5401 tttttgtttt gtacactaag attctatttc ggaactctta cccatgtcat tatttacttt
     5461 attttcatta ttgtaatttt ttaagctttt attatgaatt atttaaatac aattcttttt
     5521 gatcatgtga cagacatcac tgaaagatgt gctctgcgtt attagtcaga ttgaaatata
     5581 cattttcatg tagatatgat gttcttactt taaatgttgt tccatttgct agcttaaata
     5641 gacgatatta taaatttaag tattcactaa aaatctgacc cattaatgtt agaattagta
     5701 ttattttttg tgatattaca tttttatatt ttggatttca accaatgctc tgcgatatga
     5761 ttcaacagaa taatatgtac aagctacttt aattcatata tactaataga ttcacagatc
     5821 cagtacgact gcaagaattt acgttttgtt tgaaatggta gtagctacac taaattaaca
     5881 atgatcgatt aatccacatc cacagtgtgt aatcatccta atgaaaacaa gaataactac
     5941 aaaaatgaat ataaactttt tgcatgttag tctgacgtct aagaaaccat tattatcatg
     6001 acattaacct ataaaaatag gcgtatcacg aggccctttc gtc
//
