LOCUS       sequence-63458-e        5839 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 5839)
  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 5839)
  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-HHT2
FEATURES             Location/Qualifiers
     source          1..5839
                     /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     2517..2533
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2517..2533
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2567..2583)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2568..2584)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(3020..3036)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3020..3036)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3033..3055)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3044..3060
                     /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(3068..3074,3075..3092,3093..3098))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3113..3134
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3251..3268)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3422..4010)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3502..3521)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4181..4972,4973..5041))
                     /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     4804..4823
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5042..5146)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5812..5830
                     /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 gctctgttat tgattattta tttatttgtt
     1681 gttattttat tgtatttttt tttaaaaagc tactaataca caacaaaatg taaaccacaa
     1741 actctcccaa acagaacgaa caccctggct ttatatacaa aatttcttat cccgaaatga
     1801 ttgtcaacaa gataaacaaa aagaagccac cacaggaacc ctaccccccc tctctcccac
     1861 ccccatcgtg tcttcccttc caccttccac ccttcacctg gccatagtaa cttgaggtgt
     1921 gcacactcca gaacggtcga cccactacgg atctgaattc cagtgcgaag ttgcgggaac
     1981 ggatttatta tgaccaggcg gctactaaga agctcaaacc atcaaaaacg cgcgaagaaa
     2041 ctggtctacc cgtagcaagt tactattctc caaattagtg tttgtcgtgt gccgtgcgtc
     2101 gtgggtagac cgagattttc gcgcgcctca ggtggttcga gggcaaataa ccatagttaa
     2161 gccgttccca ttctttctca catgtacgtg ccggacgctt cgatgaaatt tttgtgaatc
     2221 gtttcgttgc ttctgtgacc acaaaacagc ctggagagga ggcgttgcgt aagggggggg
     2281 ggcctgatga gaaacagatg acacaacgca gaactggctg cagtgcaaag gcatttcctc
     2341 tgccacacac actaccataa ccagtctcgg tattaactgg ctcaacaaac gataagagcc
     2401 aaagaggaag aaaaaaaggt atttaaagga tgtatttctt tatctcctga ttagatttct
     2461 tacttgtaca gaaactgtaa ctaaaaaaaa caacaacaaa acacaacaca tacatatcta
     2521 gaactagtgg atcccccggg ctgcaggaat tcgatatcaa gcttatcgat accgtcgacc
     2581 tcgagaacac aacccacagc taccaccatc aacaatattt atatatataa cgtacacata
     2641 gaaatcacac aaacagagta tttattctta actacatgaa ctaccatcag accgtctggg
     2701 ccactatata atgtgccatt cataaacgtg atcactttac gtagcaggca accccaggtg
     2761 aaaatttttc agcgagctgc cagattgtca ggtgaaaaac tgaaaaaaac ttctgggcga
     2821 tgagcttgtg gcgggaaatt aagtatataa agcagttagt ttcttctgct tcttgtggtt
     2881 ctgggtattc ttgtaatatt tggaagaata gaaaatcaag actactgcct ttcttttcat
     2941 attatagagg aagaaatacg cacgaacacg atatagaggt aaaggtacct tgcaggcatg
     3001 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     3061 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     3121 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     3181 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     3241 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3301 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3361 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3421 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3481 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3541 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3601 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3661 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3721 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3781 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3841 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3901 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3961 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4021 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4081 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4141 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4201 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4261 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4321 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4381 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4441 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4501 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4561 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4621 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4681 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4741 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4801 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4861 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4921 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4981 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5041 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5101 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5161 aagtgccacc tgacgtcggg atctctaatg atgcaatttt tgttttgtac actaagattc
     5221 tatttcggaa ctcttaccca tgtcattatt tactttattt tcattattgt aattttttaa
     5281 gcttttatta tgaattattt aaatacaatt ctttttgatc atgtgacaga catcactgaa
     5341 agatgtgctc tgcgttatta gtcagattga aatatacatt ttcatgtaga tatgatgttc
     5401 ttactttaaa tgttgttcca tttgctagct taaatagacg atattataaa tttaagtatt
     5461 cactaaaaat ctgacccatt aatgttagaa ttagtattat tttttgtgat attacatttt
     5521 tatattttgg atttcaacca atgctctgcg atatgattca acagaataat atgtacaagc
     5581 tactttaatt catatatact aatagattca cagatccagt acgactgcaa gaatttacgt
     5641 tttgtttgaa atggtagtag ctacactaaa ttaacaatga tcgattaatc cacatccaca
     5701 gtgtgtaatc atcctaatga aaacaagaat aactacaaaa atgaatataa actttttgca
     5761 tgttagtctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     5821 atcacgaggc cctttcgtc
//
