LOCUS       40924_45678        6705 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway vector pUGW61 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6705)
  AUTHORS   Tanaka Y, Nakagawa T.
  TITLE     Gateway Vectors for transient expression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6705)
  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 6705)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6705)
  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            "Unpublished"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..6705
                     /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..1821
                     /codon_start=1
                     /label=TagRFP
                     /note="monomeric derivative of red fluorescent protein from
                     Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     protein_bind    1831..1955
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1992..2022
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             2076..2732
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             3077..3379
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(3423..3547)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3573..3825
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3834..3850)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4063..4518
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4800..4904
                     /label=AmpR promoter
     CDS             4905..5762
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5936..6524
                     /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 atggtgtcta agggcgaaga gctgattaag
     1141 gagaacatgc acatgaagct gtacatggag ggcaccgtga acaaccacca cttcaagtgc
     1201 acatccgagg gcgaaggcaa gccctacgag ggcacccaga ccatgagaat caaggtggtc
     1261 gagggcggcc ctctcccctt cgccttcgac atcctggcta ccagcttcat gtacggcagc
     1321 agaaccttca tcaaccacac ccagggcatc cccgacttct ttaagcagtc cttccctgag
     1381 ggcttcacat gggagagagt caccacatac gaagacgggg gcgtgctgac cgctacccag
     1441 gacaccagcc tccaggacgg ctgcctcatc tacaacgtca agatcagagg ggtgaacttc
     1501 ccatccaacg gccctgtgat gcagaagaaa acactcggct gggaggccaa caccgagatg
     1561 ctgtaccccg ctgacggcgg cctggaaggc agaagcgaca tggccctgaa gctcgtgggc
     1621 gggggccacc tgatctgcaa cttcaagacc acatacagat ccaagaaacc cgctaagaac
     1681 ctcaagatgc ccggcgtcta ctatgtggac cacagactgg aaagaatcaa ggaggccgac
     1741 aaagagacct acgtcgagca gcacgaggtg gctgtggcca gatactgcga cctccctagc
     1801 aaactggggc acaaacttaa tgctgttatc acaagtttgt acaaaaaagc tgaacgagaa
     1861 acgtaaaatg atataaatat caatatatta aattagattt tgcataaaaa acagactaca
     1921 taatactgta aaacacaaca tatccagtca ctatggcggc ccctcgaggg gccgcattag
     1981 gcaccccagg ctttacactt tatgcttccg gctcgtataa tgtgtggatt ttgagttagg
     2041 atccgtcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata
     2101 ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg
     2161 ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa
     2221 agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg
     2281 ctcatccgga attccgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc
     2341 acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat
     2401 accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg
     2461 aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc
     2521 cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc
     2581 ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga
     2641 ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt aatgaattac
     2701 aacagtactg cgatgagtgg cagggcgggg cgtaaacgcg tggatccggc ttactaaaag
     2761 ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg tataagaata tatactgata
     2821 tgtatacccg aagtatgtca aaaagaggta tgctatgaag cagcgtatta cagtgacagt
     2881 tgacagcgac agctatcagt tgctcaaggc atatatgatg tcaatatctc cggtctggta
     2941 agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat
     3001 caggaaggga tggctgaggt cgcccggttt attgaaatga acggctcttt tgctgacgag
     3061 aacaggggct ggtgaaatgc agtttaaggt ttacacctat aaaagagaga gccgttatcg
     3121 tctgtttgtg gatgtacaga gtgatattat tgacacgccc gggcgacgga tggtgatccc
     3181 cctggccagt gcacgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca
     3241 tatcggggat gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtctccgt
     3301 tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa
     3361 cctgatgttc tggggaatat aaatgtcagg ctcccttata cacagccagt ctgcaggtcg
     3421 accatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa
     3481 tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcagct ttcttgtaca
     3541 aagtggtgat aacctagagc tcgaatttcc ccgatcgttc aaacatttgg caataaagtt
     3601 tcttaagatt gaatcctgtt gccggtcttg cgatgattat catataattt ctgttgaatt
     3661 acgttaagca tgtaataatt aacatgtaat gcatgacgtt atttatgaga tgggttttta
     3721 tgattagagt cccgcaatta tacatttaat acgcgataga aaacaaaata tagcgcgcaa
     3781 actaggataa attatcgcgc gcggtgtcat ctatgttact agatcgggaa ttcactggcc
     3841 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     3901 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     3961 caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat
     4021 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
     4081 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4141 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4201 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4261 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4321 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4381 gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     4441 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4501 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     4561 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     4621 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     4681 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     4741 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     4801 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4861 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     4921 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4981 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     5041 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     5101 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     5161 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     5221 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5281 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5341 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5401 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5461 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5521 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5581 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5641 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5701 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5761 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     5821 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5881 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5941 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6001 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6061 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     6121 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6181 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6241 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6301 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6361 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6421 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6481 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6541 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6601 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     6661 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaag
//
