LOCUS       sequence-63303-e        4530 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 4530)
  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 4530)
  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: pBM16
FEATURES             Location/Qualifiers
     source          1..4530
                     /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"
     CDS             complement(1323..1715)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIRPQIKAFER
                     PKTFSSKVFSIMLHAYTRLYRKKRKI"
     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"
     misc_feature    1642..1698
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1711..1727)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1711..1727)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1724..1746)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1735..1751
                     /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(1759..1765,1766..1783,1784..1789))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1804..1825
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1942..1959)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2113..2701)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2193..2212)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2872..3663,3664..3732))
                     /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     3495..3514
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3733..3837)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4503..4521
                     /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 ctcggtaccc ggggatcctc tagagtcgac
     1681 ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     1741 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     1801 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     1861 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     1921 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     1981 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2041 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2101 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2161 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2221 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2281 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2341 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2401 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2461 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2521 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     2581 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2641 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2701 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     2761 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     2821 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     2881 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     2941 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3001 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3061 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3121 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3181 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3241 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3301 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3361 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     3421 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     3481 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     3541 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     3601 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     3661 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     3721 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     3781 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     3841 atttccccga aaagtgccac ctgacgtcgg gatctctaat gatgcaattt ttgttttgta
     3901 cactaagatt ctatttcgga actcttaccc atgtcattat ttactttatt ttcattattg
     3961 taatttttta agcttttatt atgaattatt taaatacaat tctttttgat catgtgacag
     4021 acatcactga aagatgtgct ctgcgttatt agtcagattg aaatatacat tttcatgtag
     4081 atatgatgtt cttactttaa atgttgttcc atttgctagc ttaaatagac gatattataa
     4141 atttaagtat tcactaaaaa tctgacccat taatgttaga attagtatta ttttttgtga
     4201 tattacattt ttatattttg gatttcaacc aatgctctgc gatatgattc aacagaataa
     4261 tatgtacaag ctactttaat tcatatatac taatagattc acagatccag tacgactgca
     4321 agaatttacg ttttgtttga aatggtagta gctacactaa attaacaatg atcgattaat
     4381 ccacatccac agtgtgtaat catcctaatg aaaacaagaa taactacaaa aatgaatata
     4441 aactttttgc atgttagtct gacgtctaag aaaccattat tatcatgaca ttaacctata
     4501 aaaataggcg tatcacgagg ccctttcgtc
//
