LOCUS       40924_45583        6672 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway vector pUGW24 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6672)
  AUTHORS   Tanaka Y, Nakagawa T.
  TITLE     Gateway Vectors for transient expression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6672)
  AUTHORS   Tanaka Y, Nakagawa T.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-APR-2011) Contact:Tsuyoshi Nakagawa Shimane 
            University, Center for Integrated Research in Science; 1060 
            Nishikawatsu, Matsue, Shimane 690-8504, Japan
REFERENCE   3  (bases 1 to 6672)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6672)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (19-APR-2011) Contact:Tsuyoshi Nakagawa Shimane University, Center 
            for Integrated Research in Science; 1060 Nishikawatsu, Matsue, 
            Shimane 690-8504, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     constructed using pUC119.
FEATURES             Location/Qualifiers
     source          1..6672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        745..1090
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             1111..1764
                     /codon_start=1
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             1771..1788
                     /codon_start=1
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
                     /translation="LVPRGS"
     protein_bind    1798..1922
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1959..1989
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             2043..2699
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             3044..3346
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(3390..3514)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3540..3792
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3801..3817)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4030..4485
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4767..4871
                     /label=AmpR promoter
     CDS             4872..5729
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5903..6491
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg
      241 catgcctgca ggtccccaga ttagcctttt caatttcaga aagaatgcta acccacagat
      301 ggttagagag gcttacgcag caggtctcat caagacgatc tacccgagca ataatctcca
      361 ggaaatcaaa taccttccca agaaggttaa agatgcagtc aaaagattca ggactaactg
      421 catcaagaac acagagaaag atatatttct caagatcaga agtactattc cagtatggac
      481 gattcaaggc ttgcttcaca aaccaaggca agtaatagag attggagtct ctaaaaaggt
      541 agttcccact gaatcaaagg ccatggagtc aaagattcaa atagaggacc taacagaact
      601 cgccgtaaag actggcgaac agttcataca gagtctctta cgactcaatg acaagaagaa
      661 aatcttcgtc aacatggtgg agcacgacac acttgtctac tccaaaaata tcaaagatac
      721 agtctcagaa gaccaaaggg caattgagac ttttcaacaa agggtaatat ccggaaacct
      781 cctcggattc cattgcccag ctatctgtca ctttattgtg aagatagtgg aaaaggaagg
      841 tggctcctac aaatgccatc attgcgataa aggaaaggcc atcgttgaag atgcctctgc
      901 cgacagtggt cccaaagatg gacccccacc cacgaggagc atcgtggaaa aagaagacgt
      961 tccaaccacg tcttcaaagc aagtggattg atgtgatatc tccactgacg taagggatga
     1021 cgcacaatcc cactatcctt cgcaagaccc ttcctctata taaggaagtt catttcattt
     1081 ggagagaaca cgggggactc tagaatgagc atgtccccta tactaggtta ttggaaaatt
     1141 aagggccttg tgcaacccac tcgacttctt ttggaatatc ttgaagaaaa atatgaagag
     1201 catttgtatg agcgcgatga aggtgataaa tggcgaaaca aaaagtttga attgggtttg
     1261 gagtttccca atcttcctta ttatattgat ggtgatgtta aattaacaca gtctatggcc
     1321 atcatacgtt atatagctga caagcacaac atgttgggtg gttgtccaaa agagcgtgca
     1381 gagatttcaa tgcttgaagg agcggttttg gatattagat acggtgtttc gagaattgca
     1441 tatagtaaag actttgaaac tctcaaagtt gattttctta gcaagctacc tgaaatgctg
     1501 aaaatgttcg aagatcgttt atgtcataaa acatatttaa atggtgatca tgtaacccat
     1561 cctgacttca tgttgtatga cgctcttgat gttgttttat acatggaccc aatgtgcctg
     1621 gatgcgttcc caaaattagt ttgttttaaa aaacgtattg aagctatccc acaaattgat
     1681 aagtacttga aatccagcaa gtatatagca tggcctttgc agggctggca agccacgttt
     1741 ggtggtggcg accatcctcc aaaatcggat ctggttccgc gtggatccgc tgttatcaca
     1801 agtttgtaca aaaaagctga acgagaaacg taaaatgata taaatatcaa tatattaaat
     1861 tagattttgc ataaaaaaca gactacataa tactgtaaaa cacaacatat ccagtcacta
     1921 tggcggcccc tcgaggggcc gcattaggca ccccaggctt tacactttat gcttccggct
     1981 cgtataatgt gtggattttg agttaggatc cgtcgagatt ttcaggagct aaggaagcta
     2041 aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag
     2101 aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg
     2161 atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta
     2221 ttcacattct tgcccgcctg atgaatgctc atccggaatt ccgtatggca atgaaagacg
     2281 gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg
     2341 aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat
     2401 attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg
     2461 agaatatgtt tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg
     2521 tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag
     2581 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc
     2641 atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt
     2701 aaacgcgtgg atccggctta ctaaaagcca gataacagta tgcgtatttg cgcgctgatt
     2761 tttgcggtat aagaatatat actgatatgt atacccgaag tatgtcaaaa agaggtatgc
     2821 tatgaagcag cgtattacag tgacagttga cagcgacagc tatcagttgc tcaaggcata
     2881 tatgatgtca atatctccgg tctggtaagc acaaccatgc agaatgaagc ccgtcgtctg
     2941 cgtgccgaac gctggaaagc ggaaaatcag gaagggatgg ctgaggtcgc ccggtttatt
     3001 gaaatgaacg gctcttttgc tgacgagaac aggggctggt gaaatgcagt ttaaggttta
     3061 cacctataaa agagagagcc gttatcgtct gtttgtggat gtacagagtg atattattga
     3121 cacgcccggg cgacggatgg tgatccccct ggccagtgca cgtctgctgt cagataaagt
     3181 ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca tgatgaccac
     3241 cgatatggcc agtgtgccgg tctccgttat cggggaagaa gtggctgatc tcagccaccg
     3301 cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataaa tgtcaggctc
     3361 ccttatacac agccagtctg caggtcgacc atagtgactg gatatgttgt gttttacagt
     3421 attatgtagt ctgtttttta tgcaaaatct aatttaatat attgatattt atatcatttt
     3481 acgtttctcg ttcagctttc ttgtacaaag tggtgataac ctagagctcg aatttccccg
     3541 atcgttcaaa catttggcaa taaagtttct taagattgaa tcctgttgcc ggtcttgcga
     3601 tgattatcat ataatttctg ttgaattacg ttaagcatgt aataattaac atgtaatgca
     3661 tgacgttatt tatgagatgg gtttttatga ttagagtccc gcaattatac atttaatacg
     3721 cgatagaaaa caaaatatag cgcgcaaact aggataaatt atcgcgcgcg gtgtcatcta
     3781 tgttactaga tcgggaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     3841 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     3901 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
     3961 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     4021 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4081 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4141 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4201 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     4261 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4321 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt
     4381 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     4441 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     4501 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     4561 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     4621 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     4681 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     4741 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4801 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4861 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4921 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4981 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5041 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5101 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5161 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5221 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5281 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5341 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5401 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5461 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5521 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5581 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5641 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5701 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5761 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5821 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5881 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5941 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6001 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     6061 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6121 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6181 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6241 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6301 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6361 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6421 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6481 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6541 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     6601 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     6661 aggaagcgga ag
//
