LOCUS       40924_45673        6724 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway vector pUGW60 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6724)
  AUTHORS   Tanaka Y, Nakagawa T.
  TITLE     Gateway Vectors for transient expression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6724)
  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 6724)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6724)
  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            "Unpublished"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..6724
                     /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        763..1108
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     protein_bind    1130..1254
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1291..1321
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1375..2031
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2376..2678
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2722..2846)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2857..3567
                     /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"
     terminator      3592..3844
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3853..3869)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4082..4537
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4819..4923
                     /label=AmpR promoter
     CDS             4924..5781
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5955..6543
                     /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 ggtctagacc caagcttgca tgcctgcagg tccccagatt agccttttca atttcagaaa
      301 gaatgctaac ccacagatgg ttagagaggc ttacgcagca ggtctcatca agacgatcta
      361 cccgagcaat aatctccagg aaatcaaata ccttcccaag aaggttaaag atgcagtcaa
      421 aagattcagg actaactgca tcaagaacac agagaaagat atatttctca agatcagaag
      481 tactattcca gtatggacga ttcaaggctt gcttcacaaa ccaaggcaag taatagagat
      541 tggagtctct aaaaaggtag ttcccactga atcaaaggcc atggagtcaa agattcaaat
      601 agaggaccta acagaactcg ccgtaaagac tggcgaacag ttcatacaga gtctcttacg
      661 actcaatgac aagaagaaaa tcttcgtcaa catggtggag cacgacacac ttgtctactc
      721 caaaaatatc aaagatacag tctcagaaga ccaaagggca attgagactt ttcaacaaag
      781 ggtaatatcc ggaaacctcc tcggattcca ttgcccagct atctgtcact ttattgtgaa
      841 gatagtggaa aaggaaggtg gctcctacaa atgccatcat tgcgataaag gaaaggccat
      901 cgttgaagat gcctctgccg acagtggtcc caaagatgga cccccaccca cgaggagcat
      961 cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa gtggattgat gtgatatctc
     1021 cactgacgta agggatgacg cacaatccca ctatccttcg caagaccctt cctctatata
     1081 aggaagttca tttcatttgg agagaacacg ggggactcta gagttatcaa caagtttgta
     1141 caaaaaagct gaacgagaaa cgtaaaatga tataaatatc aatatattaa attagatttt
     1201 gcataaaaaa cagactacat aatactgtaa aacacaacat atccagtcac tatggcggcc
     1261 cctcgagggg ccgcattagg caccccaggc tttacacttt atgcttccgg ctcgtataat
     1321 gtgtggattt tgagttagga tccgtcgaga ttttcaggag ctaaggaagc taaaatggag
     1381 aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
     1441 gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg
     1501 gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt
     1561 cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga cggtgagctg
     1621 gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt
     1681 tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa
     1741 gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg
     1801 tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat
     1861 atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag
     1921 gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc
     1981 agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc gtaaagatct
     2041 ggatccggct tactaaaagc cagataacag tatgcgtatt tgcgcgctga tttttgcggt
     2101 ataagaatat atactgatat gtatacccga agtatgtcaa aaagaggtat gctatgaagc
     2161 agcgtattac agtgacagtt gacagcgaca gctatcagtt gctcaaggca tatatgatgt
     2221 caatatctcc ggtctggtaa gcacaaccat gcagaatgaa gcccgtcgtc tgcgtgccga
     2281 acgctggaaa gcggaaaatc aggaagggat ggctgaggtc gcccggttta ttgaaatgaa
     2341 cggctctttt gctgacgaga acaggggctg gtgaaatgca gtttaaggtt tacacctata
     2401 aaagagagag ccgttatcgt ctgtttgtgg atgtacagag tgatattatt gacacgcccg
     2461 ggcgacggat ggtgatcccc ctggccagtg cacgtctgct gtcagataaa gtctcccgtg
     2521 aactttaccc ggtggtgcat atcggggatg aaagctggcg catgatgacc accgatatgg
     2581 ccagtgtgcc ggtctccgtt atcggggaag aagtggctga tctcagccac cgcgaaaatg
     2641 acatcaaaaa cgccattaac ctgatgttct ggggaatata aatgtcaggc tcccttatac
     2701 acagccagtc tgcaggtcga ccatagtgac tggatatgtt gtgttttaca gtattatgta
     2761 gtctgttttt tatgcaaaat ctaatttaat atattgatat ttatatcatt ttacgtttct
     2821 cgttcagctt tcttgtacaa agtggttgat aacagcatgg tgtctaaggg cgaagagctg
     2881 attaaggaga acatgcacat gaagctgtac atggagggca ccgtgaacaa ccaccacttc
     2941 aagtgcacat ccgagggcga aggcaagccc tacgagggca cccagaccat gagaatcaag
     3001 gtggtcgagg gcggccctct ccccttcgcc ttcgacatcc tggctaccag cttcatgtac
     3061 ggcagcagaa ccttcatcaa ccacacccag ggcatccccg acttctttaa gcagtccttc
     3121 cctgagggct tcacatggga gagagtcacc acatacgaag acgggggcgt gctgaccgct
     3181 acccaggaca ccagcctcca ggacggctgc ctcatctaca acgtcaagat cagaggggtg
     3241 aacttcccat ccaacggccc tgtgatgcag aagaaaacac tcggctggga ggccaacacc
     3301 gagatgctgt accccgctga cggcggcctg gaaggcagaa gcgacatggc cctgaagctc
     3361 gtgggcgggg gccacctgat ctgcaacttc aagaccacat acagatccaa gaaacccgct
     3421 aagaacctca agatgcccgg cgtctactat gtggaccaca gactggaaag aatcaaggag
     3481 gccgacaaag agacctacgt cgagcagcac gaggtggctg tggccagata ctgcgacctc
     3541 cctagcaaac tggggcacaa acttaattga gcttagagct cgaatttccc cgatcgttca
     3601 aacatttggc aataaagttt cttaagattg aatcctgttg ccggtcttgc gatgattatc
     3661 atataatttc tgttgaatta cgttaagcat gtaataatta acatgtaatg catgacgtta
     3721 tttatgagat gggtttttat gattagagtc ccgcaattat acatttaata cgcgatagaa
     3781 aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg cggtgtcatc tatgttacta
     3841 gatcgggaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     3901 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     3961 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg
     4021 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
     4081 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4141 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4201 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4261 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     4321 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4381 ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta
     4441 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4501 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
     4561 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     4621 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     4681 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     4741 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     4801 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4861 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4921 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4981 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5041 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5101 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5161 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5221 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5281 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5341 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5401 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5461 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5521 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5581 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5641 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5701 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5761 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     5821 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5881 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5941 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6001 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6061 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     6121 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6181 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6241 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6301 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6361 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6421 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6481 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6541 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6601 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     6661 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     6721 gaag
//
