LOCUS       sequence-63479-e        6134 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 6134)
  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 6134)
  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
FEATURES             Location/Qualifiers
     source          1..6134
                     /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     complement(2862..2878)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2863..2879)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(3315..3331)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3315..3331)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3328..3350)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3339..3355
                     /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(3363..3369,3370..3387,3388..3393))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3408..3429
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3546..3563)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3717..4305)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3797..3816)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4476..5267,5268..5336))
                     /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     5099..5118
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5337..5441)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6107..6125
                     /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 gctcagaaag cgaagaagat atatctagtt
     1681 tttctccgca ctcttaccca ttctcatttt ttttaactca gcattttact atagattttt
     1741 tttctttctt ataatcaagt acatagaata tatgcacata gaaacaaaaa agataggcaa
     1801 gttattattc agacgcatta tcataaaaaa ctttatttat ctgttaacga aacaaaattt
     1861 agcgctattt ctcaaaggat tgtagcagca tcatgagaaa aatatcgtca atcggagcga
     1921 agtttattac gtctacacag gtaacaaggg tatctgtgta ttgaacctcg gattgaaccg
     1981 gcatattctc acacaaatga gaagcttgaa atccccacgg atcctgccac acccccaaca
     2041 attaaagtga ctaagaaaca actctagacc cttgttactt atttcactca aacttgtacg
     2101 tgccctgcta acacagttta cagataggag tccatatgac tgcaattcca cacacctcga
     2161 atggctctta tcctctcccc tctccaatag aagagacttg agctaagaaa aaatttacat
     2221 cggaattttt acagctgtgt tggtttcatc actactttgc aagcctagtt tttaccactt
     2281 cctcatatgg ttatgactct tttcctcatc tttttggtgc tttcatttct ttttgcaacg
     2341 gtttataatt tttaccttcg caattttctg attttttttt gcaaaaatat tttcacaatt
     2401 tacaaaatgc actaaaagtc cgtagagaag tgggatcagt tattattcct tcacttacta
     2461 acatcgatac atttcgaatc agctatttca tagtggttta tgctattcta atcacacatt
     2521 ttgttacaac acttccttca atacacgatt gtagttgttg tacgtatcca tatagtcgac
     2581 tcatttgcgt cgccttctct tgcttttagc aacagtcgtt ctagtttttg caagcttctg
     2641 actaattcct aaaggcaagt ataaatacat gccgtttctt gctgaaacca tatgttatct
     2701 cgtatctatg tttcattaca catttttgta acagctattt caactggtga aatactacaa
     2761 gtaaaaatat attctaccta aaatattttt gctgaacaaa gaatattaaa gactagaact
     2821 agtggatccc ccgggctgca ggaattcgat atcaagctta tcgataccgt cgacctcgag
     2881 aacacaaccc acagctacca ccatcaacaa tatttatata tataacgtac acatagaaat
     2941 cacacaaaca gagtatttat tcttaactac atgaactacc atcagaccgt ctgggccact
     3001 atataatgtg ccattcataa acgtgatcac tttacgtagc aggcaacccc aggtgaaaat
     3061 ttttcagcga gctgccagat tgtcaggtga aaaactgaaa aaaacttctg ggcgatgagc
     3121 ttgtggcggg aaattaagta tataaagcag ttagtttctt ctgcttcttg tggttctggg
     3181 tattcttgta atatttggaa gaatagaaaa tcaagactac tgcctttctt ttcatattat
     3241 agaggaagaa atacgcacga acacgatata gaggtaaagg taccttgcag gcatgcaagc
     3301 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     3361 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3421 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     3481 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3541 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3601 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3661 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3721 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3781 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3841 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3901 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3961 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4021 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4081 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4141 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4201 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4261 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4321 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4381 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4441 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4501 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4561 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4621 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4681 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4741 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4801 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4861 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4921 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4981 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5041 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5101 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5161 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5221 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5281 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5341 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5401 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5461 ccacctgacg tcgggatctc taatgatgca atttttgttt tgtacactaa gattctattt
     5521 cggaactctt acccatgtca ttatttactt tattttcatt attgtaattt tttaagcttt
     5581 tattatgaat tatttaaata caattctttt tgatcatgtg acagacatca ctgaaagatg
     5641 tgctctgcgt tattagtcag attgaaatat acattttcat gtagatatga tgttcttact
     5701 ttaaatgttg ttccatttgc tagcttaaat agacgatatt ataaatttaa gtattcacta
     5761 aaaatctgac ccattaatgt tagaattagt attatttttt gtgatattac atttttatat
     5821 tttggatttc aaccaatgct ctgcgatatg attcaacaga ataatatgta caagctactt
     5881 taattcatat atactaatag attcacagat ccagtacgac tgcaagaatt tacgttttgt
     5941 ttgaaatggt agtagctaca ctaaattaac aatgatcgat taatccacat ccacagtgtg
     6001 taatcatcct aatgaaaaca agaataacta caaaaatgaa tataaacttt ttgcatgtta
     6061 gtctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     6121 gaggcccttt cgtc
//
