LOCUS       40924_22548        4910 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pGRB2.0, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4910)
  AUTHORS   Zordan RE, Ren Y, Pan SJ, Rotondo G, Penas Ade L, Iluore J, Cormack 
            BP.
  TITLE     Expression Plasmids for Use in Candida glabrata
  JOURNAL   G3 (Bethesda) 3 (10), 1675-1686 (2013)
  PUBMED    23934995
REFERENCE   2  (bases 1 to 4910)
  AUTHORS   Zordan RE, Cormack BP.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-MAY-2013) Molecular Biology and Genetics, Johns 
            Hopkins School of Medicine, 725 N Wolfe St., Hunterian 609, 
            Baltimore, MD 21205, USA
REFERENCE   3  (bases 1 to 4910)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4910)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "G3 
            (Bethesda)"; date: "2013"; volume: "3"; issue: "10"; pages: 
            "1675-1686"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (14-MAY-2013) Molecular Biology and Genetics, Johns Hopkins School 
            of Medicine, 725 N Wolfe St., Hunterian 609, Baltimore, MD 21205, 
            USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4910
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(66..664)
                     /label=Candida glabrata CEN/ARS
                     /note="Candida glabrata CEN/ARS"
     promoter        666..737
                     /label=AmpR promoter
     CDS             738..1595
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1769..2357
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2645..2666
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2681..2711
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2719..2735
                     /label=lac operator
                     /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)."
     primer_bind     2743..2759
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2780..2798
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    2811..2918
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(2927..2945)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2955..2971)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3112..3567
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(3701..4501)
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(4502..4717)
                     /label=URA3 promoter
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagcaggt tagactaaca tgcaaaaagt ttatattcat ttttgtagtt attcttgttt
      121 tcattaggat gattacacac tgtggatgtg gattaatcga tcattgttaa tttagtgtag
      181 ctactaccat ttcaaacaaa acgtaaattc ttgcagtcgt actggatctg tgaatctatt
      241 agtatatatg aattaaagta gcttgtacat attattctgt tgaatcatat cgcagagcat
      301 tggttgaaat ccaaaatata aaaatgtaat atcacaaaaa ataatactaa ttctaacatt
      361 aatgggtcag atttttagtg aatacttaaa tttataatat cgtctattta agctagcaaa
      421 tggaacaaca tttaaagtaa gaacatcata tctacatgaa aatgtatatt tcaatctgac
      481 taataacgca gagcacatct ttcagtgatg tctgtcacat gatcaaaaag aattgtattt
      541 aaataattca taataaaagc ttaaaaaatt acaataatga aaataaagta aataatgaca
      601 tgggtaagag ttccgaaata gaatcttagt gtacaaaaca aaaattgcat cattagagat
      661 ccccacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
      721 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
      781 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
      841 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
      901 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
      961 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     1021 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     1081 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     1141 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     1201 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     1261 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     1321 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     1381 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     1441 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     1501 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     1561 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     1621 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     1681 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     1741 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     1801 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     1861 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     1921 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     1981 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2041 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2101 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     2161 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     2221 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     2281 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     2341 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     2401 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     2461 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     2521 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     2581 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     2641 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     2701 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     2761 atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg gagctccacc
     2821 gcggtggcgg ccgctctaga actagtggat cccccgggct gcaggaattc gatatcaagc
     2881 ttatcgatac cgtcgacctc gagggggggc ccggtaccca attcgcccta tagtgagtcg
     2941 tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     3001 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     3061 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct
     3121 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3181 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3241 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3301 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     3361 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3421 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     3481 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3541 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     3601 cggtatttca caccgcatag ggtaataact gatataatta aattgaagct ctaatttgtg
     3661 agtttagtat acatgcattt acttataata cagtttttta gttttgctgg ccgcatcttc
     3721 tcaaatatgc ttcccagcct gcttttctgt aacgttcacc ctctacctta gcatcccttc
     3781 cctttgcaaa tagtcctctt ccaacaataa taatgtcaga tcctgtagag accacatcat
     3841 ccacggttct atactgttga cccaatgcgt ctcccttgtc atctaaaccc acaccgggtg
     3901 tcataatcaa ccaatcgtaa ccttcatctc ttccacccat gtctctttga gcaataaagc
     3961 cgataacaaa atctttgtcg ctcttcgcaa tgtcaacagt acccttagta tattctccag
     4021 tagataggga gcccttgcat gacaattctg ctaacatcaa aaggcctcta ggttcctttg
     4081 ttacttcttc tgccgcctgc ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg
     4141 cattcgtaat gtctgcccat tctgctattc tgtatacacc cgcagagtac tgcaatttga
     4201 ctgtattacc aatgtcagca aattttctgt cttcgaagag taaaaaattg tacttggcgg
     4261 ataatgcctt tagcggctta actgtgccct ccatggaaaa atcagtcaag atatccacat
     4321 gtgtttttag taaacaaatt ttgggaccta atgcttcaac taactccagt aattccttgg
     4381 tggtacgaac atccaatgaa gcacacaagt ttgtttgctt ttcgtgcatg atattaaata
     4441 gcttggcagc aacaggacta ggatgagtag cagcacgttc cttatatgta gctttcgaca
     4501 tgatttatct tcgtttcctg caggtttttg ttctgtgcag ttgggttaag aatactgggc
     4561 aatttcatgt ttcttcaaca ctacatatgc gtatatatac caatctaagt ctgtgctcct
     4621 tccttcgttc ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa ggaaaccgaa
     4681 atcaaaaaaa agaataaaaa aaaaatgatg aattgaaaag gtggtatggt gcactctcag
     4741 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     4801 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     4861 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
//
