LOCUS       sequence-63413-e        5321 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 5321)
  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 5321)
  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-EGD2
FEATURES             Location/Qualifiers
     source          1..5321
                     /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     1999..2015
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1999..2015
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2049..2065)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2050..2066)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2502..2518)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2502..2518)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2515..2537)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2526..2542
                     /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(2550..2556,2557..2574,2575..2580))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2595..2616
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2733..2750)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2904..3492)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2984..3003)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3663..4454,4455..4523))
                     /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     4286..4305
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4524..4628)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5294..5312
                     /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 gctctgtcca cttcactcac cagtaataag
     1681 ttgcccagtc accatccctt atatacgctg tggctaccga gatggccgaa cccactccaa
     1741 atatatggga tactcaaact gaaagatgat gatgagctta ttatggtaat gctgtacgta
     1801 cattaccccc gtggcaagaa acacgtacaa cgctacattt ctcgtcgacg cttttttttt
     1861 tcaaaatttc tttcatcgaa gagctttgga gggtctcata gcacatataa gttgttattg
     1921 ctacacaatt ttggtcaatt gagtagctgt tgcaataggg acaaatcaaa aacacgtcaa
     1981 taatacagat atacaaagtc tagaactagt ggatcccccg ggctgcagga attcgatatc
     2041 aagcttatcg ataccgtcga cctcgagaac acaacccaca gctaccacca tcaacaatat
     2101 ttatatatat aacgtacaca tagaaatcac acaaacagag tatttattct taactacatg
     2161 aactaccatc agaccgtctg ggccactata taatgtgcca ttcataaacg tgatcacttt
     2221 acgtagcagg caaccccagg tgaaaatttt tcagcgagct gccagattgt caggtgaaaa
     2281 actgaaaaaa acttctgggc gatgagcttg tggcgggaaa ttaagtatat aaagcagtta
     2341 gtttcttctg cttcttgtgg ttctgggtat tcttgtaata tttggaagaa tagaaaatca
     2401 agactactgc ctttcttttc atattataga ggaagaaata cgcacgaaca cgatatagag
     2461 gtaaaggtac cttgcaggca tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg
     2521 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     2581 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     2641 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     2701 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     2761 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     2821 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     2881 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     2941 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3001 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3061 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3121 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3181 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3241 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3301 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     3361 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3421 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3481 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     3541 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     3601 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     3661 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     3721 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     3781 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     3841 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     3901 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     3961 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4021 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4081 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     4141 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4201 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     4261 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     4321 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     4381 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     4441 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     4501 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     4561 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     4621 gttccgcgca catttccccg aaaagtgcca cctgacgtcg ggatctctaa tgatgcaatt
     4681 tttgttttgt acactaagat tctatttcgg aactcttacc catgtcatta tttactttat
     4741 tttcattatt gtaatttttt aagcttttat tatgaattat ttaaatacaa ttctttttga
     4801 tcatgtgaca gacatcactg aaagatgtgc tctgcgttat tagtcagatt gaaatataca
     4861 ttttcatgta gatatgatgt tcttacttta aatgttgttc catttgctag cttaaataga
     4921 cgatattata aatttaagta ttcactaaaa atctgaccca ttaatgttag aattagtatt
     4981 attttttgtg atattacatt tttatatttt ggatttcaac caatgctctg cgatatgatt
     5041 caacagaata atatgtacaa gctactttaa ttcatatata ctaatagatt cacagatcca
     5101 gtacgactgc aagaatttac gttttgtttg aaatggtagt agctacacta aattaacaat
     5161 gatcgattaa tccacatcca cagtgtgtaa tcatcctaat gaaaacaaga ataactacaa
     5221 aaatgaatat aaactttttg catgttagtc tgacgtctaa gaaaccatta ttatcatgac
     5281 attaacctat aaaaataggc gtatcacgag gccctttcgt c
//
