LOCUS       40924_14485        6924 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pDEST416TEFt7, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6924)
  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 6924)
  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 6924)
  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 6924)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 6924)
  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 JX901382.1.
FEATURES             Location/Qualifiers
     source          1..6924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        196..416
                     /label=URA3 promoter
     CDS             417..1217
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(1351..1806)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     1951..1967
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1977..1995
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(2049..2096)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2094..2113
                     /label=T7 reverse primer
                     /note="T7 reverse primer"
     protein_bind    2163..2287
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2331..2633)
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(2978..3634)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(3688..3718)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    complement(3743..3867)
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3893..4090
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(4150..4168)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4189..4205)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4213..4229)
                     /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(4237..4267)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4282..4303)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4591..5179)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5353..6210)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6211..6315)
                     /label=AmpR promoter
     misc_feature    6352..6855
                     /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 accataccac agcttttcaa ttcaattcat catttttttt ttattctttt ttttgatttc
      241 ggtttctttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
      301 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
      361 cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt
      421 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
      481 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
      541 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
      601 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
      661 ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca
      721 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
      781 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
      841 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
      901 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
      961 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
     1021 ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga accgtggatg
     1081 atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
     1141 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
     1201 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
     1261 aaattagagc ttcaatttaa ttatatcagt tattacccta tgcggtgtga aataccgcac
     1321 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat
     1381 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     1441 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     1501 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     1561 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     1621 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     1681 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     1741 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     1801 gcgcgtcgcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     1861 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     1921 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat
     1981 acgactcact atagggcgaa ttgggtaccg ggccccccct cgaatccgga tatagttcct
     2041 cctttcagca aaaaacccct caagacccgt ttagaggccc caaggggtta tgctagttat
     2101 tgctcagcgg tggcagcagc caactcagct tcctttcggg ctttgttagc agcctcgaat
     2161 caaccacttt gtacaagaaa gctgaacgag aaacgtaaaa tgatataaat atcaatatat
     2221 taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa catatccagt
     2281 cactatggtc gacctgcaga ctggctgtgt ataagggagc ctgacattta tattccccag
     2341 aacatcaggt taatggcgtt tttgatgtca ttttcgcggt ggctgagatc agccacttct
     2401 tccccgataa cggagaccgg cacactggcc atatcggtgg tcatcatgcg ccagctttca
     2461 tccccgatat gcaccaccgg gtaaagttca cgggagactt tatctgacag cagacgtgca
     2521 ctggccaggg ggatcaccat ccgtcgcccg ggcgtgtcaa taatatcact ctgtacatcc
     2581 acaaacagac gataacggct ctctctttta taggtgtaaa ccttaaactg catttcacca
     2641 gcccctgttc tcgtcagcaa aagagccgtt catttcaata aaccgggcga cctcagccat
     2701 cccttcctga ttttccgctt tccagcgttc ggcacgcaga cgacgggctt cattctgcat
     2761 ggttgtgctt accagaccgg agatattgac atcatatatg ccttgagcaa ctgatagctg
     2821 tcgctgtcaa ctgtcactgt aatacgctgc ttcatagcat acctcttttt gacatacttc
     2881 gggtatacat atcagtatat attcttatac cgcaaaaatc agcgcgcaaa tacgcatact
     2941 gttatctggc ttttagtaag ccggatccag atctttacgc cccgccctgc cactcatcgc
     3001 agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat
     3061 gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg
     3121 tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac
     3181 tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg
     3241 ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat
     3301 cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt
     3361 aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc atacggaatt
     3421 ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct
     3481 tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg
     3541 tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat
     3601 caacggtggt atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc
     3661 tcgacggatc ctaactcaaa atccacacat tatacgagcc ggaagcataa agtgtaaagc
     3721 ctggggtgcc taatgcggcc gccatagtga ctggatatgt tgtgttttac agtattatgt
     3781 agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat tttacgtttc
     3841 tcgttcagct tttttgtaca aacttgtcct agttctagag cggccgctca gtactgacaa
     3901 taaaaagatt cttgttttca agaacttgtc atttgtatag tttttttata ttgtagttgt
     3961 tctattttaa tcaaatgtta gcgtgattta tatttttttt cgcctcgaca tcatctgccc
     4021 agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt
     4081 cgctatactg ctgtcgattc gatactaacg ccgccatcca gtgtcgaaaa cgagctccag
     4141 cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt
     4201 tcctgtgtga aattgttatc cgctcacaat tccacacaac ataggagccg gaagcataaa
     4261 gtgtaaagcc tggggtgcct aatgagtgag gtaactcaca ttaattgcgt tgcgctcact
     4321 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4381 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4441 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4501 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4561 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4621 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4681 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4741 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4801 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4861 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4921 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4981 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     5041 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5101 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5161 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5221 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5281 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5341 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5401 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5461 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5521 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5581 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5641 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5701 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5761 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5821 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5881 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5941 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6001 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6061 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6121 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6181 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6241 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6301 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gggtcctttt catcacgtgc
     6361 tataaaaata attataattt aaatttttta atataaatat ataaattaaa aatagaaagt
     6421 aaaaaaagaa attaaagaaa aaatagtttt tgttttccga agatgtaaaa gactctaggg
     6481 ggatcgccaa caaatactac cttttatctt gctcttcctg ctctcaggta ttaatgccga
     6541 attgtttcat cttgtctgtg tagaagacca cacacgaaaa tcctgtgatt ttacatttta
     6601 cttatcgtta atcgaatgta tatctattta atctgctttt cttgtctaat aaatatatat
     6661 gtaaagtacg ctttttgttg aaatttttta aacctttgtt tatttttttt tcttcattcc
     6721 gtaactcttc taccttcttt atttactttc taaaatccaa atacaaaaca taaaaataaa
     6781 taaacacaga gtaaattccc aaattattcc atcattaaaa gatacgaggc gcgtgtaagt
     6841 tacaggcaag cgatccgtcc taagaaacca ttattatcat gacattaacc tataaaaata
     6901 ggcgtatcac gaggcccttt cgtc
//
