LOCUS       40924_14470        6817 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pDEST414CYC1t7, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6817)
  AUTHORS   Giuraniuc CV, Macpherson M, Saka Y.
  TITLE     Gateway Vectors for Efficient Artificial Gene Assembly In Vitro and 
            Expression in Yeast Saccharomyces cerevisiae
  JOURNAL   PLoS ONE 8 (5), E64419 (2013)
  PUBMED    23675537
REFERENCE   2  (bases 1 to 6817)
  AUTHORS   Giuraniuc C, Saka Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-OCT-2012) School of Medical Sciences, Institute of 
            Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen 
            AB25 2ZD, UK
REFERENCE   3  (bases 1 to 6817)
  AUTHORS   Giuraniuc C, Saka Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-JUL-2015) School of Medical Sciences, Institute of 
            Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen 
            AB25 2ZD, UK
REFERENCE   4  (bases 1 to 6817)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 6817)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; 
            date: "2013"; volume: "8"; issue: "5"; pages: "E64419"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (04-OCT-2012) School of Medical Sciences, Institute of Medical 
            Sciences, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, 
            UK"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (28-JUL-2015) School of Medical Sciences, Institute of Medical 
            Sciences, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, 
            UK"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     On Jul 28, 2015 this sequence version replaced JX901384.1.
FEATURES             Location/Qualifiers
     source          1..6817
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        187..467
                     /label=TRP1 promoter
     CDS             468..1139
                     /codon_start=1
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(1241..1696)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     1841..1857
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1867..1885
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(1939..1986)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1984..2003
                     /label=T7 reverse primer
                     /note="T7 reverse primer"
     protein_bind    2053..2177
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2221..2523)
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(2868..3524)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(3578..3608)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    complement(3633..3757)
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3779..4025
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(4043..4061)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4082..4098)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4106..4122)
                     /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)."
     promoter        complement(4130..4160)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4175..4196)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4484..5072)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5246..6103)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6104..6208)
                     /label=AmpR promoter
     misc_feature    6245..6748
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataaacg acattactat atatataata taggaagcat ttaatagaca gcatcgtaat
      241 atatgtgtac tttgcagtta tgacgccaga tggcagtagt ggaagatatt ctttattgaa
      301 aaatagcttg tcaccttacg tacaatcttg atccggagct tttctttttt tgccgattaa
      361 gaattaattc ggtcgaaaaa agaaaaggag agggccaaga gggagggcat tggtgactat
      421 tgagcacgtg agtatacgtg attaagcaca caaaggcagc ttggagtatg tctgttatta
      481 atttcacagg tagttctggt ccattggtga aagtttgcgg cttgcagagc acagaggccg
      541 cagaatgtgc tctagattcc gatgctgact tgctgggtat tatatgtgtg cccaatagaa
      601 agagaacaat tgacccggtt attgcaagga aaatttcaag tcttgtaaaa gcatataaaa
      661 atagttcagg cactccgaaa tacttggttg gcgtgtttcg taatcaacct aaggaggatg
      721 ttttggctct ggtcaatgat tacggcattg atatcgtcca actgcatgga gatgagtcgt
      781 ggcaagaata ccaagagttc ctcggtttgc cagttattaa aagactcgta tttccaaaag
      841 actgcaacat actactcagt gcagcttcac agaaacctca ttcgtttatt cccttgtttg
      901 attcagaagc aggtgggaca ggtgaacttt tggattggaa ctcgatttct gactgggttg
      961 gaaggcaaga gagccccgaa agcttacatt ttatgttagc tggtggactg acgccagaaa
     1021 atgttggtga tgcgcttaga ttaaatggcg ttattggtgt tgatgtaagc ggaggtgtgg
     1081 agacaaatgg tgtaaaagac tctaacaaaa tagcaaattt cgtcaaaaat gctaagaaat
     1141 aggttattac tgagtagtat ttatttaagt attgtttgtg cacttgccta tgcggtgtga
     1201 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt
     1261 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     1321 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     1381 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     1441 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     1501 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     1561 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     1621 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     1681 ccgctacagg gcgcgtcgcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     1741 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     1801 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag
     1861 cgcgcgtaat acgactcact atagggcgaa ttgggtaccg ggccccccct cgaatccgga
     1921 tatagttcct cctttcagca aaaaacccct caagacccgt ttagaggccc caaggggtta
     1981 tgctagttat tgctcagcgg tggcagcagc caactcagct tcctttcggg ctttgttagc
     2041 agcctcgaat caaccacttt gtacaagaaa gctgaacgag aaacgtaaaa tgatataaat
     2101 atcaatatat taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa
     2161 catatccagt cactatggtc gacctgcaga ctggctgtgt ataagggagc ctgacattta
     2221 tattccccag aacatcaggt taatggcgtt tttgatgtca ttttcgcggt ggctgagatc
     2281 agccacttct tccccgataa cggagaccgg cacactggcc atatcggtgg tcatcatgcg
     2341 ccagctttca tccccgatat gcaccaccgg gtaaagttca cgggagactt tatctgacag
     2401 cagacgtgca ctggccaggg ggatcaccat ccgtcgcccg ggcgtgtcaa taatatcact
     2461 ctgtacatcc acaaacagac gataacggct ctctctttta taggtgtaaa ccttaaactg
     2521 catttcacca gcccctgttc tcgtcagcaa aagagccgtt catttcaata aaccgggcga
     2581 cctcagccat cccttcctga ttttccgctt tccagcgttc ggcacgcaga cgacgggctt
     2641 cattctgcat ggttgtgctt accagaccgg agatattgac atcatatatg ccttgagcaa
     2701 ctgatagctg tcgctgtcaa ctgtcactgt aatacgctgc ttcatagcat acctcttttt
     2761 gacatacttc gggtatacat atcagtatat attcttatac cgcaaaaatc agcgcgcaaa
     2821 tacgcatact gttatctggc ttttagtaag ccggatccag atctttacgc cccgccctgc
     2881 cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa
     2941 acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat
     3001 ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa
     3061 ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta
     3121 gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac
     3181 tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg
     3241 aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc
     3301 atacggaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa
     3361 aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc
     3421 tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat
     3481 tgggatatat caacggtggt atatccagtg atttttttct ccattttagc ttccttagct
     3541 cctgaaaatc tcgacggatc ctaactcaaa atccacacat tatacgagcc ggaagcataa
     3601 agtgtaaagc ctggggtgcc taatgcggcc gccatagtga ctggatatgt tgtgttttac
     3661 agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat
     3721 tttacgtttc tcgttcagct tttttgtaca aacttgtcct agttctagag cggccgcatc
     3781 atgtaattag ttatgtcacg cttacattca cgccctcccc ccacatccgc tctaaccgaa
     3841 aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttat agttatgtta
     3901 gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacag acgcgtgtac
     3961 gcatgtaaca ttatactgaa aaccttgctt gagaaggttt tgggacgctc gaaggcttta
     4021 atttgagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
     4081 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacataggag
     4141 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gaggtaactc acattaattg
     4201 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4261 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4321 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4381 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4441 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4501 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4561 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4621 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4681 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4741 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4801 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4861 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4921 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4981 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5041 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5101 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5161 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5221 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5281 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5341 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5401 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5461 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5521 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5581 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5641 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5701 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5761 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5821 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5881 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5941 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6001 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6061 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6121 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6181 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgggtcct
     6241 tttcatcacg tgctataaaa ataattataa tttaaatttt ttaatataaa tatataaatt
     6301 aaaaatagaa agtaaaaaaa gaaattaaag aaaaaatagt ttttgttttc cgaagatgta
     6361 aaagactcta gggggatcgc caacaaatac taccttttat cttgctcttc ctgctctcag
     6421 gtattaatgc cgaattgttt catcttgtct gtgtagaaga ccacacacga aaatcctgtg
     6481 attttacatt ttacttatcg ttaatcgaat gtatatctat ttaatctgct tttcttgtct
     6541 aataaatata tatgtaaagt acgctttttg ttgaaatttt ttaaaccttt gtttattttt
     6601 ttttcttcat tccgtaactc ttctaccttc tttatttact ttctaaaatc caaatacaaa
     6661 acataaaaat aaataaacac agagtaaatt cccaaattat tccatcatta aaagatacga
     6721 ggcgcgtgta agttacaggc aagcgatccg tcctaagaaa ccattattat catgacatta
     6781 acctataaaa ataggcgtat cacgaggccc tttcgtc
//
