LOCUS       sequence-63485-e        6003 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 6003)
  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 6003)
  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
FEATURES             Location/Qualifiers
     source          1..6003
                     /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     2681..2697
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2681..2697
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2731..2747)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2732..2748)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(3184..3200)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3184..3200)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3197..3219)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3208..3224
                     /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(3232..3238,3239..3256,3257..3262))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3277..3298
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3415..3432)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3586..4174)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3666..3685)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4345..5136,5137..5205))
                     /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     4968..4987
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5206..5310)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5976..5994
                     /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 gctcatacca gttacaatta gtattacaat
     1681 ggtttaccaa gtacaaatta tgtaatatgg ttataattgg cttatatgta tggaggtagg
     1741 gaactgcagg ttattgttct gttaaactac cagtaatgca agacggctaa gtcatatgac
     1801 tgccactttg tcaatcctga agaaaatgac aatagaaagt gatataatac acgtgatatg
     1861 cgtaggatgg gacccaaaga tagccacatc acatgatctt caaaaacccc gtcaaaaagg
     1921 aataaatcga aggaaaaaaa atgccacgtg actttgatag tcaaaatcag gttatattac
     1981 tatacgatcc cccacacata acctttacca cacaccgcac gggctggact tatcctatta
     2041 gaaagcggtg cagccagggc caagaaaacg cgaacgacgc agaaaaaacg tcaagcaaaa
     2101 aaactgtggt gttatttaaa gcatacaatt tcctgcctcc ctttaatgct acgggggatc
     2161 tagcaaatgg gaaaatatca catgactttg gctagaaggg gtgggaaaac agatatttta
     2221 gtcacattgt ttgatttcac gtactacacg atacactaca ctacacaata cactgcaata
     2281 cactacactg tatggttccc tccggccctg gatatgctag cgaaggatcc caagcccatc
     2341 gaggaaatca ttcaagagca tctgcaggtc tcaaatacac taagtccaaa cacaaacagc
     2401 actgagctaa tccaagacac caattgcatg ccttctcaat tagaacagtt tattacacag
     2461 cttataactg tgcatcttgc cattcttttc gagatagcca tttgcatttg attaacatgc
     2521 ttagctcgtt ctcaagccac agtaaaatgg attgcctttt gagtttccat cgttgtataa
     2581 ataggcactc attcctatag ccttgctctt tggtcttgtc tctgtaatgg taatagctgt
     2641 aagtcaggat ataacactcc agaaaagaaa cacctaacaa tctagaacta gtggatcccc
     2701 cgggctgcag gaattcgata tcaagcttat cgataccgtc gacctcgaga acacaaccca
     2761 cagctaccac catcaacaat atttatatat ataacgtaca catagaaatc acacaaacag
     2821 agtatttatt cttaactaca tgaactacca tcagaccgtc tgggccacta tataatgtgc
     2881 cattcataaa cgtgatcact ttacgtagca ggcaacccca ggtgaaaatt tttcagcgag
     2941 ctgccagatt gtcaggtgaa aaactgaaaa aaacttctgg gcgatgagct tgtggcggga
     3001 aattaagtat ataaagcagt tagtttcttc tgcttcttgt ggttctgggt attcttgtaa
     3061 tatttggaag aatagaaaat caagactact gcctttcttt tcatattata gaggaagaaa
     3121 tacgcacgaa cacgatatag aggtaaaggt accttgcagg catgcaagct tggcgtaatc
     3181 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     3241 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     3301 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3361 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     3421 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3481 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3541 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3601 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3661 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3721 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3781 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3841 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3901 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3961 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4021 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4081 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4141 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4201 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4261 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4321 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4381 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4441 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4501 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4561 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4621 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4681 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4741 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4801 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4861 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4921 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4981 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5041 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5101 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5161 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5221 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     5281 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     5341 cgggatctct aatgatgcaa tttttgtttt gtacactaag attctatttc ggaactctta
     5401 cccatgtcat tatttacttt attttcatta ttgtaatttt ttaagctttt attatgaatt
     5461 atttaaatac aattcttttt gatcatgtga cagacatcac tgaaagatgt gctctgcgtt
     5521 attagtcaga ttgaaatata cattttcatg tagatatgat gttcttactt taaatgttgt
     5581 tccatttgct agcttaaata gacgatatta taaatttaag tattcactaa aaatctgacc
     5641 cattaatgtt agaattagta ttattttttg tgatattaca tttttatatt ttggatttca
     5701 accaatgctc tgcgatatga ttcaacagaa taatatgtac aagctacttt aattcatata
     5761 tactaataga ttcacagatc cagtacgact gcaagaattt acgttttgtt tgaaatggta
     5821 gtagctacac taaattaaca atgatcgatt aatccacatc cacagtgtgt aatcatccta
     5881 atgaaaacaa gaataactac aaaaatgaat ataaactttt tgcatgttag tctgacgtct
     5941 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     6001 gtc
//
