LOCUS       40924_6122        9004 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBC214, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9004)
  AUTHORS   Frieman MB, McCaffery JM, Cormack BP.
  TITLE     Modular domain structure in the Candida glabrata adhesin Epa1p, a 
            beta1,6 glucan-cross-linked cell wall protein
  JOURNAL   Mol. Microbiol. 46 (2), 479-492 (2002)
  PUBMED    12406223
REFERENCE   2  (bases 1 to 9004)
  AUTHORS   Cormack BP, Frieman MB.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-JAN-2002) Molecular Biology and Genetics, Johns 
            Hopkins University, 725 North Wolfe Street, Baltimore, MD 21205, USA
REFERENCE   3  (bases 1 to 9004)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9004)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol. 
            Microbiol."; date: "2002"; volume: "46"; issue: "2"; pages: 
            "479-492"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (24-JAN-2002) Molecular Biology and Genetics, Johns Hopkins 
            University, 725 North Wolfe Street, Baltimore, MD 21205, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9004
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(68..571)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     promoter        608..712
                     /label=AmpR promoter
     CDS             713..1570
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1744..2332
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2620..2641
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2656..2686
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2694..2710
                     /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     2718..2734
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2755..2773
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2792..3180
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             3298..3387
                     /codon_start=1
                     /label=3xHA
                     /note="three tandem HA epitope tags"
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     primer_bind     complement(6719..6735)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      6742..6991
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(7010..7028)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7038..7054)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      7199..7654
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(7788..8588)
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(8589..8809)
                     /label=URA3 promoter
ORIGIN
        1 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
       61 taggacggat cgcttgcctg taacttacac gcgcctcgta tcttttaatg atggaataat
      121 ttgggaattt actctgtgtt tatttatttt tatgttttgt atttggattt tagaaagtaa
      181 ataaagaagg tagaagagtt acggaatgaa gaaaaaaaaa taaacaaagg tttaaaaaat
      241 ttcaacaaaa agcgtacttt acatatatat ttattagaca agaaaagcag attaaataga
      301 tatacattcg attaacgata agtaaaatgt aaaatcacag gattttcgtg tgtggtcttc
      361 tacacagaca agatgaaaca attcggcatt aatacctgag agcaggaaga gcaagataaa
      421 aggtagtatt tgttggcgat ccccctagag tcttttacat cttcggaaaa caaaaactat
      481 tttttcttta atttcttttt ttactttcta tttttaattt atatatttat attaaaaaat
      541 ttaaattata attattttta tagcacgtga tgaaaaggac ccaggtggca cttttcgggg
      601 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
      661 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
      721 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
      781 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
      841 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
      901 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
      961 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     1021 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     1081 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     1141 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     1201 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     1261 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     1321 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     1381 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     1441 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     1501 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     1561 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     1621 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     1681 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     1741 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     1801 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     1861 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     1921 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     1981 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2041 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2101 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     2161 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     2221 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2281 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     2341 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     2401 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     2461 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     2521 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     2581 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt acctcactca
     2641 ttaggcaccc caggctttac actttatgct tccggctcct atgttgtgtg gaattgtgag
     2701 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa
     2761 ccctcactaa agggaacaaa agctggagct caatgtttct actccttttt tactcttcca
     2821 gattttctcg gactccgcgc atcgccgtac cacttcaaaa cacccaagca cagcatacta
     2881 aatttcccct ctttcttcct ctagggtgtc gttaattacc cgtactaaag gtttggaaaa
     2941 gaaaaaagag accgcctcgt ttctttttct tcgtcgaaaa aggcaataaa aatttttatc
     3001 acgtttcttt ttcttgaaaa tttttttttt tgattttttt ctctttcgat gacctcccat
     3061 tgatatttaa gttaataaac ggtcttcaat ttctcaagtt tcagtttcat ttttcttgtt
     3121 ctattacaac tttttttact tcttgctcat tagaaagaaa gcatagcaat ctaatctaag
     3181 ttttctagaa aaatgatttt aaatccagct ctatttttga ataaatgtgt atgcatttac
     3241 actactctca ttttgctcct attaacgaat ggtggttatg ctacctcgac aatcttttac
     3301 ccatacgatg ttcctgacta tgcgggctat ccctatgacg tcccggacta tgcaggatcc
     3361 tatccatatg acgttccaga ttacgctgct cagtgctcga ccaatgatat cagtttagca
     3421 tcaaaagacc ccacaacctt tcctctaggg tgttcaccag atataacaac acccaaaaaa
     3481 ggactatcaa tggaacttta ttcctatgat tttcggaaga agggttcgta cccatgttgg
     3541 gatgcagcat atttagatcc caattaccct agaactggct ataaaagcca cagacttcta
     3601 gcaaaagttg atggtgttac cggcaatata aatttctatt atcatgcaac taaaggatgc
     3661 acaccccaat taggtcactt acccgcaagc tataattacc ctaagccatt aacaatgacg
     3721 aattttacta tgttacttta tggttacttc agaccaaaag taactggctt ccatacattt
     3781 accatatctg ctgatgatct attatttgta aattttggag ctggcaatgc ttttgattgc
     3841 tgcagaaggg attcaagcgc agatcatttc ggaaattatc aagcctacgc catctggggc
     3901 tcaaaaacag ctaaagatga actaacagta catttagatg cgggcgtata ttaccctatt
     3961 aggttatttt ataataacag agaatatgat ggtgcactca gttttacatt taaaacagaa
     4021 tcaaacgaaa acactgttag tgacttttca gagtattttt tctctctcga tgacacagag
     4081 gagggctgcc caggtttaat cagctacgat tcttcttgtg cttctgttaa aacatctaaa
     4141 ataattggta ttgattatca tacagagact ccaaatgaaa acttagttcc aataaccaaa
     4201 acaatttatc atcttggaat accatgtact ggtactacta ctacacccct atgtggctct
     4261 gggttttacg atccactagc aaataagtgt gtaacaataa acacatcatc tacttcatct
     4321 gtgacaaaaa caacttcaca tacaacttcc aaagaagttt cgtttcattc aagtatcagt
     4381 tcgcagaaga ctctaattcc caagagtata cctagcccat acggaccaat aaaatcacag
     4441 tctataccaa cagaaatgga aacatcatca gaaatatcat caagcgagta tgctttctca
     4501 gatgtgatta gtacaccctc ccattcgcca tacacgaaga aacatagttc gttgaatagt
     4561 agcagttata catcaactgt tatacacagt cttacatcgt attctatctc tcagggtatt
     4621 ttctccacta gccttagtga gcaaaatatt acatcgaagt cttctacaga taaattttca
     4681 actgcaacat caatgagcaa taatattaca caatcaagta ttataatatc ccaattctca
     4741 acaaacaatg aaaactatac aacgacttca atgcatactt cctccgataa aatttcaact
     4801 gaaaccttga acaattcaat tagtacaaca accagtatat tattcagtaa ttccagtacc
     4861 atattaaaaa ataacacaac aataatatcc tcagataaag atacacatta tcatcctgtt
     4921 aatcctacaa ttgtatgttc tacaaacaaa accgaaatta tttgtgcttc aataactcaa
     4981 ccttcaatat ctaattcgaa taatcactgg agctcatctg tcctaaaatt caacagcaca
     5041 acggttcgtt caacacttcc ttcatcagca ggctctaatg aaacctctat aaacgtaccc
     5101 ttctcatcta gcaccgagag caataccagt acgagttcta cctcaacctc caacagcaaa
     5161 atggtgcgct caacacttcc ttcatcagca ggctctaatg aaacctctat aagcgtaccc
     5221 ttctcatcta gcaccgagag caataccagt acgagttcta cctcaacctc caacagcaaa
     5281 atggtgcgct caacacttcc ttcatcagca ggctctaatg aaacctctat aaacgtaccc
     5341 ttctcatcta gcaccgagag caataccagt acgagttcta cctcaacctc caacagcaaa
     5401 acggtgcgct caacaccttt ttcatcagca ggcattatca tgaccagctt atcacaaaga
     5461 aataataaat cggctagcag ttacgctagt tctaattcta aatgttacaa tactgctgat
     5521 tcatgtcgca aggtacatag caccccatca tatctactaa ctagtagtta cacatctgaa
     5581 ggtgttgatt atgactgttc gttagtctca actaaattaa aaatcaatga tactaattgc
     5641 gtgaataaca aacataccac aaaatcctgt ttgaaaactt ctgttactac aacaatacca
     5701 acacttgaaa taaaaaccgc aagaaaatcc tcctccaata ctattggcct ccattcatac
     5761 cccacttcca gtcccaacaa atcaatatca tcagcaccaa ttataggcta catttcatcc
     5821 tttaagacaa ttaaaacagc cagtccatct taccaaactt cagatctcac aactatcaca
     5881 actataacaa gcttaaacaa tccaggttca acagcagtgg agaatacaca cgaatccaac
     5941 gataaatcca gaaagactag tagtaatgat atctcttcga agcatagcgt aatcaaggaa
     6001 accaaagacg ctgtagaatc atcgaacaaa tcacatcaaa caaacacact caaatgtagt
     6061 tcaattatca tagcaagctc ttcacacaac agttatgagt ctttaggtgg aacaacatta
     6121 actttgacat taagtaaagt atattcgcca cagaataatg acaccttacc atttttggaa
     6181 atatctgaag tgaacccatc gcgtacagta cttcctgaaa gttctaaaat gatgcaatat
     6241 ttaacatcaa cagaagagag gaataaaaca gcgaggaaca caatagcaac aaacatcgtc
     6301 agtataagta cattccactt cgaaggagaa ggaaatgcta ttagaatggg atacactcaa
     6361 cttctgctta tgttgatagg cataatagtg atgaacatag ggacctaaaa ccagaaaata
     6421 taataacttc tatagtttag cattgtaaaa gctgtggtgg ctgttttgtt tcactgcaat
     6481 ctttcttgca ggcggatcac atcaccatcc gtttttaaca gtgtatattg cctaggtagc
     6541 atccatcatc aatacccttt atatcagagc tttatacaaa taatattcct ggtcacactt
     6601 gaaatgagag atattataaa aaacttcggg tggtatcata agtttcaggg tttttggttt
     6661 acatacgaag cctaatttgc cttttctatt ctcccagaga attcgatatc aagcttatcg
     6721 ataccgtcga cctcgatcga gtcatgtaat tagttatgtc acgcttacat tcacgccctc
     6781 ctcccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta
     6841 tttatttttt ttaatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct
     6901 tttttttctg tacaaacgcg tgtacgcatg taacattata ctgaaaacct tgcttgagaa
     6961 ggttttggga cgctcgaagg ctttaatttg cggccggtac ccaattcgcc ctatagtgag
     7021 tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     7081 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     7141 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcgac
     7201 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     7261 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     7321 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     7381 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     7441 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     7501 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     7561 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     7621 gcgaatttta acaaaatatt aacgtttaca atttcctgat gcggtatttt ctccttacgc
     7681 atctgtgcgg tatttcacac cgcatagggt aataactgat ataattaaat tgaagctcta
     7741 atttgtgagt ttagtataca tgcatttact tataatacag ttttttagtt ttgctggccg
     7801 catcttctca aatatgcttc ccagcctgct tttctgtaac gttcaccctc taccttagca
     7861 tcccttccct ttgcaaatag tcctcttcca acaataataa tgtcagatcc tgtagagacc
     7921 acatcatcca cggttctata ctgttgaccc aatgcgtctc ccttgtcatc taaacccaca
     7981 ccgggtgtca taatcaacca atcgtaacct tcatctcttc cacccatgtc tctttgagca
     8041 ataaagccga taacaaaatc tttgtcgctc ttcgcaatgt caacagtacc cttagtatat
     8101 tctccagtag atagggagcc cttgcatgac aattctgcta acatcaaaag gcctctaggt
     8161 tcctttgtta cttcttctgc cgcctgcttc aaaccgctaa caatacctgg gcccaccaca
     8221 ccgtgtgcat tcgtaatgtc tgcccattct gctattctgt atacacccgc agagtactgc
     8281 aatttgactg tattaccaat gtcagcaaat tttctgtctt cgaagagtaa aaaattgtac
     8341 ttggcggata atgcctttag cggcttaact gtgccctcca tggaaaaatc agtcaagata
     8401 tccacatgtg tttttagtaa acaaattttg ggacctaatg cttcaactaa ctccagtaat
     8461 tccttggtgg tacgaacatc caatgaagca cacaagtttg tttgcttttc gtgcatgata
     8521 ttaaatagct tggcagcaac aggactagga tgagtagcag cacgttcctt atatgtagct
     8581 ttcgacatga tttatcttcg tttcctgcag gtttttgttc tgtgcagttg ggttaagaat
     8641 actgggcaat ttcatgtttc ttcaacacta catatgcgta tatataccaa tctaagtctg
     8701 tgctccttcc ttcgttcttc cttctgttcg gagattaccg aatcaaaaaa atttcaaaga
     8761 aaccgaaatc aaaaaaaaga ataaaaaaaa aatgatgaat tgaattgaaa agctgtggta
     8821 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     8881 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     8941 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     9001 gcga
//
