LOCUS       40924_45543        6130 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway vector pUGW14 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6130)
  AUTHORS   Tanaka Y, Nakagawa T.
  TITLE     Gateway Vectors for transient expression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6130)
  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 6130)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6130)
  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            "Unpublished"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..6130
                     /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             2875..2964
                     /codon_start=1
                     /label=3xHA
                     /note="three tandem HA epitope tags"
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     terminator      2998..3250
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3259..3275)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3488..3943
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4225..4329
                     /label=AmpR promoter
     CDS             4330..5187
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5361..5949
                     /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 aacagcgggt taattaacat cttttaccca
     2881 tacgatgttc ctgactatgc gggctatccc tatgacgtcc cggactatgc aggatcctat
     2941 ccatatgacg ttccagatta cgctgctcag tgcagcgctt agagctcgaa tttccccgat
     3001 cgttcaaaca tttggcaata aagtttctta agattgaatc ctgttgccgg tcttgcgatg
     3061 attatcatat aatttctgtt gaattacgtt aagcatgtaa taattaacat gtaatgcatg
     3121 acgttattta tgagatgggt ttttatgatt agagtcccgc aattatacat ttaatacgcg
     3181 atagaaaaca aaatatagcg cgcaaactag gataaattat cgcgcgcggt gtcatctatg
     3241 ttactagatc gggaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     3301 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     3361 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct
     3421 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     3481 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3541 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3601 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3661 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     3721 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3781 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt
     3841 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3901 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     3961 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     4021 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     4081 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     4141 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     4201 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4261 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4321 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4381 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4441 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4501 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4561 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4621 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4681 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4741 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4801 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4861 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4921 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4981 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5041 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5101 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5161 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5221 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5281 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5341 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5401 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5461 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     5521 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5581 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5641 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5701 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5761 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5821 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5881 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5941 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6001 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     6061 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     6121 gaagcggaag
//
