LOCUS       40924_14460        6999 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pDEST413CYC1t7, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6999)
  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 6999)
  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 6999)
  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 6999)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 6999)
  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 JX901383.1.
FEATURES             Location/Qualifiers
     source          1..6999
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        317..503
                     /label=HIS3 promoter
     CDS             504..1160
                     /codon_start=1
                     /label=HIS3
                     /note="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      complement(1423..1878)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     2023..2039
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2049..2067
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(2121..2168)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2166..2185
                     /label=T7 reverse primer
                     /note="T7 reverse primer"
     protein_bind    2235..2359
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2403..2705)
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(3050..3706)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(3760..3790)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    complement(3815..3939)
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3961..4207
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(4225..4243)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4264..4280)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4288..4304)
                     /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(4312..4342)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4357..4378)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4666..5254)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5428..6285)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6286..6390)
                     /label=AmpR promoter
     misc_feature    6427..6930
                     /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 tcagcgcgtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataaatt cccgttttaa gagcttggtg agcgctagga gtcactgcca ggtatcgttt
      241 gaacacggca ttagtcaggg aagtcataac acagtccttt cccgcaattt tctttttcta
      301 ttactcttgg cctcctctag tacactctat atttttttat gcctcggtaa tgattttcat
      361 tttttttttt cccctagcgg atgactcttt ttttttctta gcgattggca ttatcacata
      421 atgaattata cattatataa agtaatgtga tttcttcgaa gaatatacta aaaaatgagc
      481 aggcaagata aacgaaggca aagatgacag agcagaaagc cctagtaaag cgtattacaa
      541 atgaaaccaa gattcagatt gcgatctctt taaagggtgg tcccctagcg atagagcact
      601 cgatcttccc agaaaaagag gcagaagcag tagcagaaca ggccacacaa tcgcaagtga
      661 ttaacgtcca cacaggtata gggtttctgg accatatgat acatgctctg gccaagcatt
      721 ccggctggtc gctaatcgtt gagtgcattg gtgacttaca catagacgac catcacacca
      781 ctgaagactg cgggattgct ctcggtcaag cttttaaaga ggccctactg gcgcgtggag
      841 taaaaaggtt tggatcagga tttgcgcctt tggatgaggc actttccaga gcggtggtag
      901 atctttcgaa caggccgtac gcagttgtcg aacttggttt gcaaagggag aaagtaggag
      961 atctctcttg cgagatgatc ccgcattttc ttgaaagctt tgcagaggct agcagaatta
     1021 ccctccacgt tgattgtctg cgaggcaaga atgatcatca ccgtagtgag agtgcgttca
     1081 aggctcttgc ggttgccata agagaagcca cctcgcccaa tggtaccaac gatgttccct
     1141 ccaccaaagg tgttcttatg tagtgacacc gattatttaa agctgcagca tacgatatat
     1201 atacatgtgt atatatgtat acctatgaat gtcagtaagt atgtatacga acagtatgat
     1261 actgaagatg acaaggtaat gcatcattct atacgtgtca ttctgaacga ggcgcgcttt
     1321 ccttttttct ttttgctttt tctttttttt tctcttgaac tcgacggatc tatgcggtgt
     1381 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta aacgttaata
     1441 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     1501 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     1561 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     1621 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     1681 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     1741 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     1801 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     1861 cgccgctaca gggcgcgtcg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
     1921 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     1981 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     2041 agcgcgcgta atacgactca ctatagggcg aattgggtac cgggcccccc ctcgaatccg
     2101 gatatagttc ctcctttcag caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt
     2161 tatgctagtt attgctcagc ggtggcagca gccaactcag cttcctttcg ggctttgtta
     2221 gcagcctcga atcaaccact ttgtacaaga aagctgaacg agaaacgtaa aatgatataa
     2281 atatcaatat attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac
     2341 aacatatcca gtcactatgg tcgacctgca gactggctgt gtataaggga gcctgacatt
     2401 tatattcccc agaacatcag gttaatggcg tttttgatgt cattttcgcg gtggctgaga
     2461 tcagccactt cttccccgat aacggagacc ggcacactgg ccatatcggt ggtcatcatg
     2521 cgccagcttt catccccgat atgcaccacc gggtaaagtt cacgggagac tttatctgac
     2581 agcagacgtg cactggccag ggggatcacc atccgtcgcc cgggcgtgtc aataatatca
     2641 ctctgtacat ccacaaacag acgataacgg ctctctcttt tataggtgta aaccttaaac
     2701 tgcatttcac cagcccctgt tctcgtcagc aaaagagccg ttcatttcaa taaaccgggc
     2761 gacctcagcc atcccttcct gattttccgc tttccagcgt tcggcacgca gacgacgggc
     2821 ttcattctgc atggttgtgc ttaccagacc ggagatattg acatcatata tgccttgagc
     2881 aactgatagc tgtcgctgtc aactgtcact gtaatacgct gcttcatagc atacctcttt
     2941 ttgacatact tcgggtatac atatcagtat atattcttat accgcaaaaa tcagcgcgca
     3001 aatacgcata ctgttatctg gcttttagta agccggatcc agatctttac gccccgccct
     3061 gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac
     3121 aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat
     3181 atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa
     3241 aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt
     3301 tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa
     3361 actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat
     3421 ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg
     3481 ccatacggaa ttccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat
     3541 aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg
     3601 tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc
     3661 attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag
     3721 ctcctgaaaa tctcgacgga tcctaactca aaatccacac attatacgag ccggaagcat
     3781 aaagtgtaaa gcctggggtg cctaatgcgg ccgccatagt gactggatat gttgtgtttt
     3841 acagtattat gtagtctgtt ttttatgcaa aatctaattt aatatattga tatttatatc
     3901 attttacgtt tctcgttcag cttttttgta caaacttgtc ctagttctag agcggccgca
     3961 tcatgtaatt agttatgtca cgcttacatt cacgccctcc ccccacatcc gctctaaccg
     4021 aaaaggaagg agttagacaa cctgaagtct aggtccctat ttattttttt atagttatgt
     4081 tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac agacgcgtgt
     4141 acgcatgtaa cattatactg aaaaccttgc ttgagaaggt tttgggacgc tcgaaggctt
     4201 taatttgagc tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     4261 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatagg
     4321 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgaggtaac tcacattaat
     4381 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4441 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4501 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4561 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4621 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4681 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4741 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4801 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4861 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4921 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4981 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5041 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5101 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5161 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5221 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5281 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5341 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5401 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5461 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5521 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5581 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5641 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5701 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5761 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5821 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5881 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5941 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6001 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6061 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6121 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6181 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6241 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6301 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6361 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgggtc
     6421 cttttcatca cgtgctataa aaataattat aatttaaatt ttttaatata aatatataaa
     6481 ttaaaaatag aaagtaaaaa aagaaattaa agaaaaaata gtttttgttt tccgaagatg
     6541 taaaagactc tagggggatc gccaacaaat actacctttt atcttgctct tcctgctctc
     6601 aggtattaat gccgaattgt ttcatcttgt ctgtgtagaa gaccacacac gaaaatcctg
     6661 tgattttaca ttttacttat cgttaatcga atgtatatct atttaatctg cttttcttgt
     6721 ctaataaata tatatgtaaa gtacgctttt tgttgaaatt ttttaaacct ttgtttattt
     6781 ttttttcttc attccgtaac tcttctacct tctttattta ctttctaaaa tccaaataca
     6841 aaacataaaa ataaataaac acagagtaaa ttcccaaatt attccatcat taaaagatac
     6901 gaggcgcgtg taagttacag gcaagcgatc cgtcctaaga aaccattatt atcatgacat
     6961 taacctataa aaataggcgt atcacgaggc cctttcgtc
//
