LOCUS       40924_9601        8010 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Gateway(R) destination vector for mammalian cell expression of 
            proteins C-terminally modified with V5 and 6xHis tags plus a 
            cleavable biotinylation peptide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8010)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8010)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        199..578
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        579..782
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        827..845
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    875..999
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1431..1733)
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(2083..2736)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVFDGFHVGRMLNELQQYCDEWQGG"
     promoter        complement(2737..2839)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     protein_bind    complement(3017..3141)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3167..3208
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     CDS             3218..3502
                     /codon_start=1
                     /label=capTEV(TM)
                     /note="composite tag for in vivo biotinylation, TEV
                     protease cleavage, and 6xHis tagging"
                     /translation="HHHHHHGENLYFQGQLENLYFQGGAGTPVTAPLAGTIWKVLASEG
                     QTVAAGEVLLILEAMKMETEIRAAQAGTVRGIAVKAGDAVAVGDTLMTLA"
     polyA_signal    3589..3637
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3839..4267
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4281..4610
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4677..5468
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5647..5780
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5817..5833)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5841..5857)
                     /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(5865..5895)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5910..5931)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6219..6807)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6981..7838)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7839..7943)
                     /label=AmpR promoter
ORIGIN
        1 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
       61 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
      121 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
      181 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
      241 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      301 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      361 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      421 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      481 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      541 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      601 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      661 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      721 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      781 ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat acgactcact
      841 atagggagac ccaagctggc tagttaagct atcaacaagt ttgtacaaaa aagctgaacg
      901 agaaacgtaa aatgatataa atatcaatat attaaattag attttgcata aaaaacagac
      961 tacataatac tgtaaaacac aacatatcca gtcactatga atcaactact tagatggtat
     1021 tagtgacctg tagtcgaccg acagccttcc aaatgttctt cgggtgatgc tgccaactta
     1081 gtcgaccgac agccttccaa atgttcttct caaacggaat cgtcgtatcc agcctactcg
     1141 ctattgtcct caatgccgta ttaaatcata aaaagaaata agaaaaagag gtgcgagcct
     1201 cttttttgtg tgacaaaata aaaacatcta cctattcata tacgctagtg tcatagtcct
     1261 gaaaatcatc tgcatcaaga acaatttcac aactcttata cttttctctt acaagtcgtt
     1321 cggcttcatc tggattttca gcctctatac ttactaaacg tgataaagtt tctgtaattt
     1381 ctactgtatc gacctgcaga ctggctgtgt ataagggagc ctgacattta tattccccag
     1441 aacatcaggt taatggcgtt tttgatgtca ttttcgcggt ggctgagatc agccacttct
     1501 tccccgataa cggagaccgg cacactggcc atatcggtgg tcatcatgcg ccagctttca
     1561 tccccgatat gcaccaccgg gtaaagttca cgggagactt tatctgacag cagacgtgca
     1621 ctggccaggg ggatcaccat ccgtcgcccg ggcgtgtcaa taatatcact ctgtacatcc
     1681 acaaacagac gataacggct ctctctttta taggtgtaaa ccttaaactg catttcacca
     1741 gcccctgttc tcgtcagcaa aagagccgtt catttcaata aaccgggcga cctcagccat
     1801 cccttcctga ttttccgctt tccagcgttc ggcacgcaga cgacgggctt cattctgcat
     1861 ggttgtgctt accagaccgg agatattgac atcatatatg ccttgagcaa ctgatagctg
     1921 tcgctgtcaa ctgtcactgt aatacgctgc ttcatagcat acctcttttt gacatacttc
     1981 gggtatacat atcagtatat attcttatac cgcaaaaatc agcgcgcaaa tacgcatact
     2041 gttatctggc ttttagtaag ccggatccac gcggcgttta cgccccccct gccactcatc
     2101 gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcaa agacggcatg
     2161 atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat
     2221 ggtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa
     2281 actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt tagggaaata
     2341 ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa
     2401 atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt
     2461 gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg ccatacggaa
     2521 ttccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg
     2581 cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata
     2641 ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat
     2701 atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa
     2761 tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga
     2821 acctcttacg tgccgatcaa cgtctcattt tcgccaaaag ttggcccagg gcttcccggt
     2881 atcaacaggg acaccaggat ttatttattc tgcgaagtga tcttccgtca caggtattta
     2941 ttcggcgcaa agtgcgtcgg gtgatgctgc caacttagtc gactacaggt cactaatacc
     3001 atctaagtag ttgattcata gtgactggat atgttgtgtt ttacagtatt atgtagtctg
     3061 ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg tttctcgttc
     3121 agctttcttg tacaaagtgg ttgatctaga gggcccgcgg ttcgaaggta agcctatccc
     3181 taaccctctc ctcggtctcg attctacgcg taccgggcat catcatcatc atcatggtga
     3241 gaatctttat tttcagggtc aattggagaa tctttatttt cagggtggcg ccggcacccc
     3301 ggtgaccgcc ccgctggcgg gcactatctg gaaggtgctg gccagcgaag gccagacggt
     3361 ggccgcaggc gaggtgctgc tgattctgga agccatgaag atggaaaccg aaatccgcgc
     3421 cgcgcaggcc gggaccgtgc gcggtatcgc ggtgaaagcc ggcgacgcgg tggcggtcgg
     3481 cgacaccctg atgaccctgg cgggctctgg atcctagtaa tgagtttaaa cgggggaggc
     3541 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     3601 gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     3661 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     3721 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     3781 ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac
     3841 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3901 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3961 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4021 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     4081 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4141 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     4201 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4261 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4321 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4381 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4441 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4501 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     4561 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     4621 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     4681 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     4741 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4801 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     4861 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4921 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4981 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     5041 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     5101 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     5161 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     5221 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     5281 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     5341 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     5401 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     5461 agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg
     5521 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5581 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5641 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5701 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5761 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     5821 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5881 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5941 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     6001 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     6061 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     6121 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6181 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6241 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6301 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6361 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6421 cgctgtatgt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6481 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6541 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6601 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     6661 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6721 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6781 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6841 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6901 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6961 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     7021 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     7081 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     7141 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     7201 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     7261 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     7321 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7381 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     7441 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7501 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7561 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7621 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7681 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7741 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7801 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7861 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7921 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     7981 tcgggagatc tcccgatccc ctatggtgca
//
