LOCUS       40924_6127        8275 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBC542, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8275)
  AUTHORS   Zupancic ML, Frieman M, Smith D, Alvarez RA, Cummings RD, Cormack 
            BP.
  TITLE     Glycan microarray analysis of Candida glabrata adhesin ligand 
            specificity
  JOURNAL   Mol. Microbiol. 68 (3), 547-559 (2008)
  PUBMED    18394144
REFERENCE   2  (bases 1 to 8275)
  AUTHORS   Cormack BP.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-JAN-2008) Molecular Biology and Genetics, Johns 
            Hopkins Medical Institutes, 725 N. Wolfe St., 613 Hunterian, 
            Baltimore, MD 21205, USA
REFERENCE   3  (bases 1 to 8275)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8275)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol. 
            Microbiol."; date: "2008"; volume: "68"; issue: "3"; pages: 
            "547-559"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (22-JAN-2008) Molecular Biology and Genetics, Johns Hopkins Medical 
            Institutes, 725 N. Wolfe St., 613 Hunterian, Baltimore, MD 21205, 
            USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8275
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(70..573)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     promoter        610..714
                     /label=AmpR promoter
     CDS             715..1572
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1746..2334
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2622..2643
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2658..2688
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2696..2712
                     /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     2720..2736
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2757..2775
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2794..3182
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     protein_bind    3195..3319
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        3344..3374
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             3428..4084
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             4429..4731
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(4775..4899)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             4915..5004
                     /codon_start=1
                     /label=3xHA
                     /note="three tandem HA epitope tags"
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     misc_feature    5020..5742
                     /label=EPA1 Ser/Thr rich region
                     /note="EPA1 Ser/Thr rich region"
     misc_feature    5743..5967
                     /label=CWP2 C-terminal domain
                     /note="CWP2 C-terminal domain"
     primer_bind     complement(5991..6007)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      6013..6262
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(6281..6299)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6309..6325)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6470..6925
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(7059..7859)
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(7860..8080)
                     /label=URA3 promoter
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata
      121 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt
      181 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa
      241 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata
      301 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct
      361 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata
      421 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact
      481 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa
      541 atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg
      601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      841 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
      901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
      961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1141 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1261 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1501 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1861 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     1921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2101 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2221 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2341 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2401 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2461 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2521 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     2581 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact
     2641 cattaggcac cccaggcttt acactttatg cttccggctc ctatgttgtg tggaattgtg
     2701 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     2761 aaccctcact aaagggaaca aaagctggag ctcaatgttt ctactccttt tttactcttc
     2821 cagattttct cggactccgc gcatcgccgt accacttcaa aacacccaag cacagcatac
     2881 taaatttccc ctctttcttc ctctagggtg tcgttaatta cccgtactaa aggtttggaa
     2941 aagaaaaaag agaccgcctc gtttcttttt cttcgtcgaa aaaggcaata aaaattttta
     3001 tcacgtttct ttttcttgaa aatttttttt tttgattttt ttctctttcg atgacctccc
     3061 attgatattt aagttaataa acggtcttca atttctcaag tttcagtttc atttttcttg
     3121 ttctattaca acttttttta cttcttgctc attagaaaga aagcatagca atctaatcta
     3181 agttttctag aatcacaagt ttgtacaaaa aagctgaacg agaaacgtaa aatgatataa
     3241 atatcaatat attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac
     3301 aacatatcca gtcactatgg cggccgcatt aggcacccca ggctttacac tttatgcttc
     3361 cggctcgtat aatgtgtgga ttttgagtta ggatccggcg agattttcag gagctaagga
     3421 agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg
     3481 taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca
     3541 gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc
     3601 ctttattcac attcttgccc gcctgatgaa tgctcatccg gaattccgta tggcaatgaa
     3661 agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca
     3721 aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca
     3781 catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt
     3841 tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt
     3901 aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac
     3961 gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg tctgtgatgg
     4021 cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg
     4081 ggcgtaaacg cgtggatccg gcttactaaa agccagataa cagtatgcgt atttgcgcgc
     4141 tgatttttgc ggtataagaa tatatactga tatgtatacc cgaagtatgt caaaaagagg
     4201 tgtgctatga agcagcgtat tacagtgaca gttgacagcg acagctatca gttgctcaag
     4261 gcatatatga tgtcaatatc tccggtctgg taagcacaac catgcagaat gaagcccgtc
     4321 gtctgcgtgc cgaacgctgg aaagcggaaa atcaggaagg gatggctgag gtcgcccggt
     4381 ttattgaaat gaacggctct tttgctgacg agaacaggga ctggtgaaat gcagtttaag
     4441 gtttacacct ataaaagaga gagccgttat cgtctgtttg tggatgtaca gagtgatatt
     4501 attgacacgc ccgggcgacg gatggtgatc cccctggcca gtgcacgtct gctgtcagat
     4561 aaagtctccc gtgaacttta cccggtggtg catatcgggg atgaaagctg gcgcatgatg
     4621 accaccgata tggccagtgt gccggtctcc gttatcgggg aagaagtggc tgatctcagc
     4681 caccgcgaaa atgacatcaa aaacgccatt aacctgatgt tctggggaat ataaatgtca
     4741 ggctccctta tacacagcca gtctgcaggt cgaccatagt gactggatat gttgtgtttt
     4801 acagtattat gtagtctgtt ttttatgcaa aatctaattt aatatattga tatttatatc
     4861 attttacgtt tctcgttcag ctttcttgta caaagtggtg gctcgagaat cttttaccca
     4921 tacgatgttc ctgactatgc gggctatccc tatgacgtcc cggactatgc aggatcctat
     4981 ccatatgacg ttccagatta cgctgctcag tggtcgagcg tgacaaaaac aacttcacat
     5041 acaacttcca aagaagtttc gtttcattca agtatcagtt cgcagaagac tctaattccc
     5101 aagagtatac ctagcccata cggaccaata aaatcacagt ctataccaac agaaatggaa
     5161 acatcatcag aaatatcatc aagcgagtat gctttctcag atgtgattag tacaccctcc
     5221 cattcgccat acacgaagaa acatagttcg ttgaatagta gcagttatac atcaactgtt
     5281 atacacagtc ttacatcgta ttctatctct cagggtattt tctccactag ccttagtgag
     5341 caaaatatta catcgaagtc ttctacagat aaattttcaa ctgcaacatc aatgagcaat
     5401 aatattacac aatcaagtat tataatatcc caattctcaa caaacaatga aaactataca
     5461 acgacttcaa tgcatacttc ctccgataaa atttcaactg aaaccttgaa caattcaatt
     5521 agtacaacaa ccagtatatt attcagtaat tccagtacca tattaaaaaa taacacaaca
     5581 ataatatcct cagataaaga tacacattat catcctgtta atcctacaat tgtatgttct
     5641 acaaacaaaa ccgaaattat ttgtgcttca ataactcaac cttcaatatc taattcgaat
     5701 aatcactgga gctcatctgt cctaaaattc aacagcacaa cgtcgagtga atccgctgcc
     5761 gccatttctc aaatcactga cggtcaaatc caagctacta ccactgctac caccgaagct
     5821 accaccactg ctgccccatc ttccaccgtt gaaactgttt ctccatccag caccgaaact
     5881 atctctcaac aaactgaaaa tggtgctgct aaggccgctg tcggtatggg tgccggtgct
     5941 ctagctgctg ctgctatgtt gttataataa gaattcgata tcaagcttat cgataccgtc
     6001 gacctcgtcg agtcatgtaa ttagttatgt cacgcttaca ttcacgccct cctcccacat
     6061 ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt
     6121 tttaatagtt atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct
     6181 gtacaaacgc gtgtacgcat gtaacattat actgaaaacc ttgcttgaga aggttttggg
     6241 acgctcgaag gctttaattt gcggccggta cccaattcgc cctatagtga gtcgtattac
     6301 gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     6361 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     6421 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcga cgcgccctgt
     6481 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     6541 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     6601 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     6661 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     6721 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     6781 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     6841 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     6901 aacaaaatat taacgtttac aatttcctga tgcggtattt tctccttacg catctgtgcg
     6961 gtatttcaca ccgcataggg taataactga tataattaaa ttgaagctct aatttgtgag
     7021 tttagtatac atgcatttac ttataataca gttttttagt tttgctggcc gcatcttctc
     7081 aaatatgctt cccagcctgc ttttctgtaa cgttcaccct ctaccttagc atcccttccc
     7141 tttgcaaata gtcctcttcc aacaataata atgtcagatc ctgtagagac cacatcatcc
     7201 acggttctat actgttgacc caatgcgtct cccttgtcat ctaaacccac accgggtgtc
     7261 ataatcaacc aatcgtaacc ttcatctctt ccacccatgt ctctttgagc aataaagccg
     7321 ataacaaaat ctttgtcgct cttcgcaatg tcaacagtac ccttagtata ttctccagta
     7381 gatagggagc ccttgcatga caattctgct aacatcaaaa ggcctctagg ttcctttgtt
     7441 acttcttctg ccgcctgctt caaaccgcta acaatacctg ggcccaccac accgtgtgca
     7501 ttcgtaatgt ctgcccattc tgctattctg tatacacccg cagagtactg caatttgact
     7561 gtattaccaa tgtcagcaaa ttttctgtct tcgaagagta aaaaattgta cttggcggat
     7621 aatgccttta gcggcttaac tgtgccctcc atggaaaaat cagtcaagat atccacatgt
     7681 gtttttagta aacaaatttt gggacctaat gcttcaacta actccagtaa ttccttggtg
     7741 gtacgaacat ccaatgaagc acacaagttt gtttgctttt cgtgcatgat attaaatagc
     7801 ttggcagcaa caggactagg atgagtagca gcacgttcct tatatgtagc tttcgacatg
     7861 atttatcttc gtttcctgca ggtttttgtt ctgtgcagtt gggttaagaa tactgggcaa
     7921 tttcatgttt cttcaacact acatatgcgt atatatacca atctaagtct gtgctccttc
     7981 cttcgttctt ccttctgttc ggagattacc gaatcaaaaa aatttcaaag aaaccgaaat
     8041 caaaaaaaag aataaaaaaa aaatgatgaa ttgaattgaa aagctgtggt atggtgcact
     8101 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     8161 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     8221 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcga
//
