LOCUS       40924_45633        6727 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway vector pUGW44 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6727)
  AUTHORS   Tanaka Y, Nakagawa T.
  TITLE     Gateway Vectors for transient expression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6727)
  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 6727)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6727)
  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            "Unpublished"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..6727
                     /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..3573
                     /codon_start=1
                     /label=ECFP
                     /note="enhanced CFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     terminator      3595..3847
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3856..3872)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4085..4540
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4822..4926
                     /label=AmpR promoter
     CDS             4927..5784
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5958..6546
                     /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 tgagcaaggg cgaggagctg
     2881 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     2941 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     3001 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctggggc
     3061 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     3121 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     3181 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     3241 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacatcagc
     3301 cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc
     3361 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     3421 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     3481 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     3541 gggatcactc tcggcatgga cgagctgtac aaggcttaga gctcgaattt ccccgatcgt
     3601 tcaaacattt ggcaataaag tttcttaaga ttgaatcctg ttgccggtct tgcgatgatt
     3661 atcatataat ttctgttgaa ttacgttaag catgtaataa ttaacatgta atgcatgacg
     3721 ttatttatga gatgggtttt tatgattaga gtcccgcaat tatacattta atacgcgata
     3781 gaaaacaaaa tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc atctatgtta
     3841 ctagatcggg aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     3901 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     3961 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat
     4021 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca
     4081 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4141 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4201 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4261 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
     4321 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4381 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat
     4441 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4501 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca
     4561 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     4621 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     4681 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     4741 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     4801 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4861 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4921 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     4981 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5041 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5101 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5161 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5221 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5281 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5341 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5401 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5461 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     5521 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     5581 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5641 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5701 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     5761 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     5821 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5881 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5941 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6001 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6061 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     6121 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6181 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6241 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6301 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6361 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6421 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6481 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6541 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6601 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     6661 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     6721 gcggaag
//
