LOCUS       40924_45643        6717 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway vector pUGW5 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6717)
  AUTHORS   Tanaka Y, Nakagawa T.
  TITLE     Gateway Vectors for transient expression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6717)
  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 6717)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6717)
  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            "Unpublished"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..6717
                     /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"
     protein_bind    1108..1232
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1269..1299
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1353..2009
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2354..2656
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2700..2824)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2841..3557
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     terminator      3585..3837
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3846..3862)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4075..4530
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4812..4916
                     /label=AmpR promoter
     CDS             4917..5774
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5948..6536
                     /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 taatcaaaca agtttgtaca aaaaagctga acgagaaacg
     1141 taaaatgata taaatatcaa tatattaaat tagattttgc ataaaaaaca gactacataa
     1201 tactgtaaaa cacaacatat ccagtcacta tggcggcccc tcgaggggcc gcattaggca
     1261 ccccaggctt tacactttat gcttccggct cgtataatgt gtggattttg agttaggatc
     1321 cgtcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca
     1381 ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc
     1441 aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga
     1501 aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc
     1561 atccggaatt ccgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc
     1621 cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc
     1681 acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa
     1741 acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtctca gccaatccct
     1801 gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg
     1861 ttttcaccat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc
     1921 aggttcatca tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac
     1981 agtactgcga tgagtggcag ggcggggcgt aatctagagg atccggctta ctaaaagcca
     2041 gataacagta tgcgtatttg cgcgctgatt tttgcggtat aagaatatat actgatatgt
     2101 atacccgaag tatgtcaaaa agaggtatgc tatgaagcag cgtattacag tgacagttga
     2161 cagcgacagc tatcagttgc tcaaggcata tatgatgtca atatctccgg tctggtaagc
     2221 acaaccatgc agaatgaagc ccgtcgtctg cgtgccgaac gctggaaagc ggaaaatcag
     2281 gaagggatgg ctgaggtcgc ccggtttatt gaaatgaacg gctcttttgc tgacgagaac
     2341 aggggctggt gaaatgcagt ttaaggttta cacctataaa agagagagcc gttatcgtct
     2401 gtttgtggat gtacagagtg atattattga cacgcccggg cgacggatgg tgatccccct
     2461 ggccagtgca cgtctgctgt cagataaagt cccccgtgaa ctttacccgg tggtgcatat
     2521 cggggatgaa agctggcgca tgatgaccac cgatatggcc agtgtgccgg tctccgttat
     2581 cggggaagaa gtggctgatc tcagccaccg cgaaaatgac atcaaaaacg ccattaacct
     2641 gatgttctgg ggaatataaa tgtcaggctc ccttatacac agccagtctg caggtcgacc
     2701 atagtgactg gatatgttgt gttttacagt attatgtagt ctgtttttta tgcaaaatct
     2761 aatttaatat attgatattt atatcatttt acgtttctcg ttcagctttc ttgtacaaag
     2821 tggttcgatc tagaggatcc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     2881 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     2941 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     3001 tgcccgtgcc ctggcccacc ctcgtgacca ccttcaccta cggcgtgcag tgcttcagcc
     3061 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     3121 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     3181 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     3241 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     3301 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     3361 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     3421 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     3481 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actcacggca
     3541 tggacgagct gtacaagtaa agcggcccga gctcgaattt ccccgatcgt tcaaacattt
     3601 ggcaataaag tttcttaaga ttgaatcctg ttgccggtct tgcgatgatt atcatataat
     3661 ttctgttgaa ttacgttaag catgtaataa ttaacatgta atgcatgacg ttatttatga
     3721 gatgggtttt tatgattaga gtcccgcaat tatacattta atacgcgata gaaaacaaaa
     3781 tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc atctatgtta ctagatcggg
     3841 aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     3901 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     3961 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt
     4021 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     4081 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4141 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4201 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     4261 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     4321 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4381 tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga
     4441 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4501 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     4561 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     4621 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     4681 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     4741 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     4801 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     4861 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     4921 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4981 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     5041 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     5101 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     5161 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     5221 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     5281 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     5341 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     5401 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     5461 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     5521 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     5581 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     5641 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     5701 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     5761 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     5821 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5881 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5941 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6001 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6061 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     6121 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6181 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6241 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6301 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6361 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6421 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6481 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6541 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     6601 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     6661 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaag
//
