LOCUS       40924_9611        7711 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Gateway(R) destination vector for high-level expression of 
            C-terminally V5-tagged proteins in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7711)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7711)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7711
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    911..1035
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1467..1769)
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(2114..2770)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(2771..2873)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     protein_bind    complement(3051..3175)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3201..3242
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    3326..3374
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3576..4004
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4018..4347
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4414..5205
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5384..5517
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5554..5570)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5578..5594)
                     /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)."
     promoter        complement(5602..5632)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5647..5668)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5956..6544)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6718..7575)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7576..7680)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagctatca acaagtttgt acaaaaaagc tgaacgagaa acgtaaaatg atataaatat
      961 caatatatta aattagattt tgcataaaaa acagactaca taatactgta aaacacaaca
     1021 tatccagtca ctatgaatca actacttaga tggtattagt gacctgtagt cgaccgacag
     1081 ccttccaaat gttcttcggg tgatgctgcc aacttagtcg accgacagcc ttccaaatgt
     1141 tcttctcaaa cggaatcgtc gtatccagcc tactcgctat tgtcctcaat gccgtattaa
     1201 atcataaaaa gaaataagaa aaagaggtgc gagcctcttt tttgtgtgac aaaataaaaa
     1261 catctaccta ttcatatacg ctagtgtcat agtcctgaaa atcatctgca tcaagaacaa
     1321 tttcacaact cttatacttt tctcttacaa gtcgttcggc ttcatctgga ttttcagcct
     1381 ctatacttac taaacgtgat aaagtttctg taatttctac tgtatcgacc tgcagactgg
     1441 ctgtgtataa gggagcctga catttatatt ccccagaaca tcaggttaat ggcgtttttg
     1501 atgtcatttt cgcggtggct gagatcagcc acttcttccc cgataacgga gaccggcaca
     1561 ctggccatat cggtggtcat catgcgccag ctttcatccc cgatatgcac caccgggtaa
     1621 agttcacggg agactttatc tgacagcaga cgtgcactgg ccagggggat caccatccgt
     1681 cgcccgggcg tgtcaataat atcactctgt acatccacaa acagacgata acggctctct
     1741 cttttatagg tgtaaacctt aaactgcatt tcaccagtcc ctgttctcgt cagcaaaaga
     1801 gccgttcatt tcaataaacc gggcgacctc agccatccct tcctgatttt ccgctttcca
     1861 gcgttcggca cgcagacgac gggcttcatt ctgcatggtt gtgcttacca gaccggagat
     1921 attgacatca tatatgcctt gagcaactga tagctgtcgc tgtcaactgt cactgtaata
     1981 cgctgcttca tagcacacct ctttttgaca tacttcgggt atacatatca gtatatattc
     2041 ttataccgca aaaatcagcg cgcaaatacg catactgtta tctggctttt agtaagccgg
     2101 atccacgcga ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca
     2161 ttctgccgac atggaagcca tcacagacgg catgatgaac ctgaatcgcc agcggcatca
     2221 gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt
     2281 ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgagacga
     2341 aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca
     2401 catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg
     2461 atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata
     2521 tcaccagctc accgtctttc attgccatac ggaattccgg atgagcattc atcaggcggg
     2581 caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa
     2641 aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg
     2701 cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt
     2761 ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg
     2821 gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaacgtctc
     2881 attttcgcca aaagttggcc cagggcttcc cggtatcaac agggacacca ggatttattt
     2941 attctgcgaa gtgatcttcc gtcacaggta tttattcggc gcaaagtgcg tcgggtgatg
     3001 ctgccaactt agtcgactac aggtcactaa taccatctaa gtagttgatt catagtgact
     3061 ggatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc taatttaata
     3121 tattgatatt tatatcattt tacgtttctc gttcagcttt cttgtacaaa gtggttgatc
     3181 tagagggccc gcggttcgaa ggtaagccta tccctaaccc tctcctcggt ctcgattcta
     3241 cgcgtaccgg ttagtaatga gtttaaacgg gggaggctaa ctgaaacacg gaaggagaca
     3301 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt
     3361 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     3421 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     3481 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     3541 agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     3601 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3661 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3721 tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3781 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3841 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3901 actcaaccct atctcggtct attcttttga tttataaggg attttgggga tttcggccta
     3961 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     4021 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4081 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4141 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4201 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4261 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     4321 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     4381 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     4441 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4501 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4561 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     4621 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4681 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4741 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4801 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4861 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4921 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     4981 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5041 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     5101 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     5161 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     5221 ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5281 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5341 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     5401 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5461 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     5521 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5581 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5641 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5701 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5761 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5821 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5881 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5941 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6001 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6061 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6121 tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
     6181 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6241 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6301 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6361 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     6421 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6481 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6541 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6601 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6661 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6721 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6781 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6841 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6901 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6961 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7021 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7081 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7141 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7201 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7261 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7321 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7381 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7441 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7501 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7561 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7621 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7681 cacatttccc cgaaaagtgc cacctgacgt c
//
