LOCUS       sequence-63468-e        5950 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 5950)
  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 5950)
  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
FEATURES             Location/Qualifiers
     source          1..5950
                     /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     2628..2644
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2628..2644
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2678..2694)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2679..2695)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(3131..3147)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3131..3147)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3144..3166)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3155..3171
                     /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(3179..3185,3186..3203,3204..3209))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3224..3245
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3362..3379)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3533..4121)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3613..3632)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4292..5083,5084..5152))
                     /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     4915..4934
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5153..5257)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5923..5941
                     /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 tgaattgaga gctcagcatt tttatacacg ttttacgtat
     1681 tttttttgca atacacatag atacgtacgt acaacccttt ctattgtgag aactttgtaa
     1741 cactcttttg tagaccagtg tagtcagcaa gcatcagcaa gcatcagcaa gcatcagcaa
     1801 gcatcagcta gtctaaagcc tttagcaaag tcaaaaacca acacccacac accgggatcc
     1861 caccttcccc gagatgcctg ttcccgactc ccagctgctc ccacgacggg gaggaatcag
     1921 aagacggaga tagaaagaga gcgttgaacg cgcgcgatgc caatgagaga taaaaaaaaa
     1981 gaaataccaa tcccacgaca tattctaacg ggctccacag acaatgcaca acaatcgaga
     2041 tgggattgaa tcacagagag agagatcaga aaaaaaagga actgcaaggg tccccacccc
     2101 aaactcccct ccgctatcca ctacaatcgc cgcaccgaca aaatccattg tgtccgcaac
     2161 gaccagagca aagagccaaa aacacaattg aataacaaag aaaagggcag agagccactt
     2221 agacaattac gtaggaacca tatttttgca taactttatg caatttgccc aaattcaagt
     2281 gaagcgagag aagccagaga gctacagctg aagagctaca gctgaagtga aaagccaaga
     2341 atctcttaga atataccgtt tggcttattt gaatcaaata tttatgcgct taaatcccat
     2401 aaaaaagcaa tttatacaga taagtctgca gaaagaaaaa ttttcttctt gatacctgaa
     2461 caaagaaatc aaactcatca agaataaatc aattcatgaa aaaaaaacat atataaagga
     2521 caacatggaa tcaagtttca ataattttta gattgtacat aaataaagag accagactaa
     2581 tacaactgta taagctctaa acattcaatt gccaaaaaac attaacatct agaactagtg
     2641 gatcccccgg gctgcaggaa ttcgatatca agcttatcga taccgtcgac ctcgagaaca
     2701 caacccacag ctaccaccat caacaatatt tatatatata acgtacacat agaaatcaca
     2761 caaacagagt atttattctt aactacatga actaccatca gaccgtctgg gccactatat
     2821 aatgtgccat tcataaacgt gatcacttta cgtagcaggc aaccccaggt gaaaattttt
     2881 cagcgagctg ccagattgtc aggtgaaaaa ctgaaaaaaa cttctgggcg atgagcttgt
     2941 ggcgggaaat taagtatata aagcagttag tttcttctgc ttcttgtggt tctgggtatt
     3001 cttgtaatat ttggaagaat agaaaatcaa gactactgcc tttcttttca tattatagag
     3061 gaagaaatac gcacgaacac gatatagagg taaaggtacc ttgcaggcat gcaagcttgg
     3121 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     3181 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     3241 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     3301 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     3361 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3421 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3481 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3541 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3601 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3661 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3721 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3781 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3841 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3901 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3961 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4021 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4081 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4141 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4201 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4261 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4321 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4381 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4441 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4501 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4561 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4621 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4681 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4741 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4801 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4861 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4921 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4981 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5041 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5101 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5161 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5221 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5281 ctgacgtcgg gatctctaat gatgcaattt ttgttttgta cactaagatt ctatttcgga
     5341 actcttaccc atgtcattat ttactttatt ttcattattg taatttttta agcttttatt
     5401 atgaattatt taaatacaat tctttttgat catgtgacag acatcactga aagatgtgct
     5461 ctgcgttatt agtcagattg aaatatacat tttcatgtag atatgatgtt cttactttaa
     5521 atgttgttcc atttgctagc ttaaatagac gatattataa atttaagtat tcactaaaaa
     5581 tctgacccat taatgttaga attagtatta ttttttgtga tattacattt ttatattttg
     5641 gatttcaacc aatgctctgc gatatgattc aacagaataa tatgtacaag ctactttaat
     5701 tcatatatac taatagattc acagatccag tacgactgca agaatttacg ttttgtttga
     5761 aatggtagta gctacactaa attaacaatg atcgattaat ccacatccac agtgtgtaat
     5821 catcctaatg aaaacaagaa taactacaaa aatgaatata aactttttgc atgttagtct
     5881 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     5941 ccctttcgtc
//
