LOCUS       sequence-63482-e        6856 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 6856)
  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 6856)
  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-LYS21-GFP
FEATURES             Location/Qualifiers
     source          1..6856
                     /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"
     CDS             2847..3563
                     /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(4037..4053)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4037..4053)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4050..4072)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4061..4077
                     /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(4085..4091,4092..4109,4110..4115))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4130..4151
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4268..4285)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4439..5027)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4519..4538)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5198..5989,5990..6058))
                     /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     5821..5840
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6059..6163)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6829..6847
                     /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 ctcagaaagc gaagaagata tatctagttt
     1681 ttctccgcac tcttacccat tctcattttt tttaactcag cattttacta tagatttttt
     1741 ttctttctta taatcaagta catagaatat atgcacatag aaacaaaaaa gataggcaag
     1801 ttattattca gacgcattat cataaaaaac tttatttatc tgttaacgaa acaaaattta
     1861 gcgctatttc tcaaaggatt gtagcagcat catgagaaaa atatcgtcaa tcggagcgaa
     1921 gtttattacg tctacacagg taacaagggt atctgtgtat tgaacctcgg attgaaccgg
     1981 catattctca cacaaatgag aagcttgaaa tccccacgga tcctgccaca cccccaacaa
     2041 ttaaagtgac taagaaacaa ctctagaccc ttgttactta tttcactcaa acttgtacgt
     2101 gccctgctaa cacagtttac agataggagt ccatatgact gcaattccac acacctcgaa
     2161 tggctcttat cctctcccct ctccaataga agagacttga gctaagaaaa aatttacatc
     2221 ggaattttta cagctgtgtt ggtttcatca ctactttgca agcctagttt ttaccacttc
     2281 ctcatatggt tatgactctt ttcctcatct ttttggtgct ttcatttctt tttgcaacgg
     2341 tttataattt ttaccttcgc aattttctga tttttttttg caaaaatatt ttcacaattt
     2401 acaaaatgca ctaaaagtcc gtagagaagt gggatcagtt attattcctt cacttactaa
     2461 catcgataca tttcgaatca gctatttcat agtggtttat gctattctaa tcacacattt
     2521 tgttacaaca cttccttcaa tacacgattg tagttgttgt acgtatccat atagtcgact
     2581 catttgcgtc gccttctctt gcttttagca acagtcgttc tagtttttgc aagcttctga
     2641 ctaattccta aaggcaagta taaatacatg ccgtttcttg ctgaaaccat atgttatctc
     2701 gtatctatgt ttcattacac atttttgtaa cagctatttc aactggtgaa atactacaag
     2761 taaaaatata ttctacctaa aatatttttg ctgaacaaag aatattaaag actagaacta
     2821 gtggatcccc cgggctgcag gaattcatgt ctaaaggtga agaattattc actggtgttg
     2881 tcccaatttt ggttgaatta gatggtgatg ttaatggtca caaattttct gtctccggtg
     2941 aaggtgaagg tgatgctact tacggtaaat tgaccttaaa atttatttgt actactggta
     3001 aattgccggt tccatggcca accttagtca ctactttcgg ttatggtgtt caatgttttg
     3061 ctagataccc agatcatatg aaacaacatg actttttcaa gtctgccatg ccagaaggtt
     3121 atgttcaaga aagaactatt tttttcaaag atgacggtaa ctacaagacc agagctgaag
     3181 tcaagtttga aggtgatacc ttagttaata gaatcgaatt aaaaggtatt gattttaaag
     3241 aagatggtaa cattttaggt cacaaattgg aatacaacta taactctcac aatgtttaca
     3301 tcatggctga caaacaaaag aatggtatca aagttaactt caaaattaga cacaacattg
     3361 aagatggttc tgttcaatta gctgaccatt atcaacaaaa tactccaatt ggtgatggtc
     3421 cagtcttgtt accagacaac cattacttat ccactcaatc tgccttatcc aaagatccaa
     3481 acgaaaagag agaccacatg gtcttgttag aatttgttac tgctgctggt attacccatg
     3541 gtatggatga attgtacaaa taactgcagg tcgacctcga gaacacaacc cacagctacc
     3601 accatcaaca atatttatat atataacgta cacatagaaa tcacacaaac agagtattta
     3661 ttcttaacta catgaactac catcagaccg tctgggccac tatataatgt gccattcata
     3721 aacgtgatca ctttacgtag caggcaaccc caggtgaaaa tttttcagcg agctgccaga
     3781 ttgtcaggtg aaaaactgaa aaaaacttct gggcgatgag cttgtggcgg gaaattaagt
     3841 atataaagca gttagtttct tctgcttctt gtggttctgg gtattcttgt aatatttgga
     3901 agaatagaaa atcaagacta ctgcctttct tttcatatta tagaggaaga aatacgcacg
     3961 aacacgatat agaggtaaag gtacccgggg atcctctaga gtcgacctgc aggcatgcaa
     4021 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     4081 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4141 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4201 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4261 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4321 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4381 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4441 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4501 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4561 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4621 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4681 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4741 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4801 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4861 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4921 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4981 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5041 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5101 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5161 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5221 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5281 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5341 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5401 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5461 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5521 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5581 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5641 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5701 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5761 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5821 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5881 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5941 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6001 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6061 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6121 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6181 tgccacctga cgtcgggatc tctaatgatg caatttttgt tttgtacact aagattctat
     6241 ttcggaactc ttacccatgt cattatttac tttattttca ttattgtaat tttttaagct
     6301 tttattatga attatttaaa tacaattctt tttgatcatg tgacagacat cactgaaaga
     6361 tgtgctctgc gttattagtc agattgaaat atacattttc atgtagatat gatgttctta
     6421 ctttaaatgt tgttccattt gctagcttaa atagacgata ttataaattt aagtattcac
     6481 taaaaatctg acccattaat gttagaatta gtattatttt ttgtgatatt acatttttat
     6541 attttggatt tcaaccaatg ctctgcgata tgattcaaca gaataatatg tacaagctac
     6601 tttaattcat atatactaat agattcacag atccagtacg actgcaagaa tttacgtttt
     6661 gtttgaaatg gtagtagcta cactaaatta acaatgatcg attaatccac atccacagtg
     6721 tgtaatcatc ctaatgaaaa caagaataac tacaaaaatg aatataaact ttttgcatgt
     6781 tagtctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     6841 acgaggccct ttcgtc
//
