LOCUS       sequence-63474-e        6972 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 6972)
  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 6972)
  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
FEATURES             Location/Qualifiers
     source          1..6972
                     /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     3650..3666
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3650..3666
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3700..3716)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3701..3717)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(4153..4169)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4153..4169)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4166..4188)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4177..4193
                     /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(4201..4207,4208..4225,4226..4231))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4246..4267
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4384..4401)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4555..5143)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4635..4654)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5314..6105,6106..6174))
                     /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     5937..5956
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6175..6279)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6945..6963
                     /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 gctcctgcag tgtcccgttt gtttcagtcc
     1681 tttgttatta tttcttgaat aatggactct cacttgggaa aggaaatact taagagtggt
     1741 attcaaacac ctagttatat agtgattccg gttttgtaat cgtctacaat ataagcaaag
     1801 tttagggaat ctagtttcat tgttcttttt tttctttctt agtgttcttt attttgccaa
     1861 ttttttttct tcttcccatt atgtggcagg aaggctcaat tttcatttag cagagctaag
     1921 aacgaaatta ctggggggac ccgcttcgtt atctcgcttt tcccttttcc tttttccttc
     1981 tgcgtaactt tctaacagac ttcttttccc tcgaggaaag attttgtgcg tcgcttctct
     2041 ccaaagttgc ttttttcatt ttttgttcag ttttcgcagt tccaagtgct acgggaatcg
     2101 tctctgccaa atgtttgtgt gtgtgctact gcgcagatcg ttagggtgga aaagaaagaa
     2161 aatgcccact gccttctccg agaaaatcgt gtgaaaaccg tgaaaaccga aatggatctt
     2221 accttttagg ctcctgcgtt gcgctctatg catcggcctg ccaattagaa atcccttcca
     2281 tacttttttt ctcttaaatc tgggagggtg gctacgcggg ttcttctgca tttgcttttt
     2341 tctatcaaaa ccaaatatca aatcttgcgg tgcttcctgc agagaagtat ctcgaggaat
     2401 agacattgtg agccaacaga aaaaaagagg gggggaggcc agagacgatg gtggatgtgc
     2461 gatgatctct ggcaccttgt taagaataga actgtaagtg ctgtgggaac tgcgcacagt
     2521 catggtgctg ggatatctaa tttcgggtcg gtggcagatc agacccacgt gggtctgatc
     2581 taaagtaaag agcagacaat aacttgatag ccagtgcttt agttccagtc acactgttaa
     2641 gcgggtaccc attgtatttt tacttgcctg gaggatgtac ggtaagtgtc aagcaaatat
     2701 tcttgagcaa aaaatttgca tgaaaacgga tccatcctct gtgaattgac acataactgg
     2761 agtagtaatt agctttctat gtgcagacgg tgagacctga cgcccattag ttagacaatg
     2821 tcgtatgggc cctctgcact atgggggggg gactatgtga ctatttattt gcgtacagta
     2881 ttacatgtat atatttcttg tgcatgtggt tcaagctgtg ttctttgaaa aatgtaactt
     2941 aataagcttc tatggccaag ttggtaaggc gccacactag taatgtggag atcatcagtt
     3001 caaatctggt tggaagcatt tattttttac tttaaaaaag catgccatta ataatcctac
     3061 tattggtgag aaaagcattc gactttttgg ttttgatact atccaggttt gtaacacagg
     3121 atactttcgt acacaccctt ttcgaataag ttttttgccc aactacacaa tcacccgtac
     3181 aatgcagggg atatccaaac cattaagtag tcatcatttt gaaaaaataa aagaaatcca
     3241 aatttccatc gggatttttc ttcagtttcc tatcaacctc tcatctttga tctaactagt
     3301 tctttacaaa aagctttcca tttcatatct tcgggaaaag ggtttccttt taggatgtgc
     3361 aagaaacatc cattggctct tgactaatac taagaattcc agtgtaaaca gtaccacctt
     3421 cccattgtca attgaagttg gaaattaaga aaaaattgcg gtgggaaata tttggcgatg
     3481 actaaacatt tattgcaagt cctacaaaat tttttttttc atagcaaact cactatatat
     3541 attaacaaca ataacttaac aaataaaaat aacaaataga agtaaaacgt ctatactgag
     3601 ccggaccttg tcttacatta caacaaacta caattcgtac ccactagcat ctagaactag
     3661 tggatccccc gggctgcagg aattcgatat caagcttatc gataccgtcg acctcgagaa
     3721 cacaacccac agctaccacc atcaacaata tttatatata taacgtacac atagaaatca
     3781 cacaaacaga gtatttattc ttaactacat gaactaccat cagaccgtct gggccactat
     3841 ataatgtgcc attcataaac gtgatcactt tacgtagcag gcaaccccag gtgaaaattt
     3901 ttcagcgagc tgccagattg tcaggtgaaa aactgaaaaa aacttctggg cgatgagctt
     3961 gtggcgggaa attaagtata taaagcagtt agtttcttct gcttcttgtg gttctgggta
     4021 ttcttgtaat atttggaaga atagaaaatc aagactactg cctttctttt catattatag
     4081 aggaagaaat acgcacgaac acgatataga ggtaaaggta ccttgcaggc atgcaagctt
     4141 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     4201 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     4261 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4321 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4381 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4441 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4501 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4561 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4621 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4681 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4741 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4801 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4861 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4921 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4981 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5041 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5101 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5161 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5221 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5281 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5341 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5401 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5461 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5521 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5581 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5641 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5701 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5761 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5821 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5881 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5941 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6001 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6061 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6121 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6181 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6241 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6301 acctgacgtc gggatctcta atgatgcaat ttttgttttg tacactaaga ttctatttcg
     6361 gaactcttac ccatgtcatt atttacttta ttttcattat tgtaattttt taagctttta
     6421 ttatgaatta tttaaataca attctttttg atcatgtgac agacatcact gaaagatgtg
     6481 ctctgcgtta ttagtcagat tgaaatatac attttcatgt agatatgatg ttcttacttt
     6541 aaatgttgtt ccatttgcta gcttaaatag acgatattat aaatttaagt attcactaaa
     6601 aatctgaccc attaatgtta gaattagtat tattttttgt gatattacat ttttatattt
     6661 tggatttcaa ccaatgctct gcgatatgat tcaacagaat aatatgtaca agctacttta
     6721 attcatatat actaatagat tcacagatcc agtacgactg caagaattta cgttttgttt
     6781 gaaatggtag tagctacact aaattaacaa tgatcgatta atccacatcc acagtgtgta
     6841 atcatcctaa tgaaaacaag aataactaca aaaatgaata taaacttttt gcatgttagt
     6901 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     6961 ggccctttcg tc
//
