LOCUS       40924_9606        7921 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Gateway(R) destination vector for mammalian cell expression of 
            proteins N-terminally modified with 6xHis and V5 tags plus a 
            cleavable biotinylation peptide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7921)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7921)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7921
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    173..193
                     /label=attB4
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     enhancer        222..601
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        602..805
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             860..1162
                     /codon_start=1
                     /label=capTEV(TM)
                     /note="composite tag for in vivo biotinylation, TEV
                     protease cleavage, and 6xHis tagging"
                     /translation="MGAGTPVTAPLAGTIWKVLASEGQTVAAGEVLLILEAMKMETEIR
                     AAQAGTVRGIAVKAGDAVAVGDTLMTLAGSGSENLYFQGQLENLYFQGQLHHHHHH"
     CDS             1163..1204
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     protein_bind    1205..1329
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1761..2063)
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(2413..3066)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG"
     promoter        complement(3067..3169)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     protein_bind    complement(3347..3471)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    3536..3584
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3786..4214
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4228..4557
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4624..5415
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5594..5727
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5764..5780)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5788..5804)
                     /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(5812..5842)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5857..5878)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6166..6754)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6928..7785)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7786..7890)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattatcgg atcaactttg
      181 tatagaaaag ttgaacgatg tacgggccag atatacgcgt tgacattgat tattgactag
      241 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      301 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      361 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      421 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      481 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      541 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      601 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      661 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      721 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      781 gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg cttactggct
      841 tatcgaaatt agcttcacca tgggcgccgg caccccggtg accgccccgc tggcgggcac
      901 tatctggaag gtgctggcca gcgaaggcca gacggtggcc gcaggcgagg tgctgctgat
      961 tctggaagcc atgaagatgg aaaccgaaat ccgcgccgcg caggccggga ccgtgcgcgg
     1021 tatcgcggtg aaagccggcg acgcggtggc ggtcggcgac accctgatga ccctggcggg
     1081 ctctggatcc gagaatcttt attttcaggg tcaattggag aatctttatt ttcagggtca
     1141 attgcatcat catcatcatc atggtaagcc tatccctaac cctctcctcg gtctcgattc
     1201 tacgacaagt ttgtacaaaa aagctgaacg agaaacgtaa aatgatataa atatcaatat
     1261 attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac aacatatcca
     1321 gtcactatga atcaactact tagatggtat tagtgacctg tagtcgaccg acagccttcc
     1381 aaatgttctt cgggtgatgc tgccaactta gtcgaccgac agccttccaa atgttcttct
     1441 caaacggaat cgtcgtatcc agcctactcg ctattgtcct caatgccgta ttaaatcata
     1501 aaaagaaata agaaaaagag gtgcgagcct cttttttgtg tgacaaaata aaaacatcta
     1561 cctattcata tacgctagtg tcatagtcct gaaaatcatc tgcatcaaga acaatttcac
     1621 aactcttata cttttctctt acaagtcgtt cggcttcatc tggattttca gcctctatac
     1681 ttactaaacg tgataaagtt tctgtaattt ctactgtatc gacctgcaga ctggctgtgt
     1741 ataagggagc ctgacattta tattccccag aacatcaggt taatggcgtt tttgatgtca
     1801 ttttcgcggt ggctgagatc agccacttct tccccgataa cggagaccgg cacactggcc
     1861 atatcggtgg tcatcatgcg ccagctttca tccccgatat gcaccaccgg gtaaagttca
     1921 cgggagactt tatctgacag cagacgtgca ctggccaggg ggatcaccat ccgtcgcccg
     1981 ggcgtgtcaa taatatcact ctgtacatcc acaaacagac gataacggct ctctctttta
     2041 taggtgtaaa ccttaaactg catttcacca gcccctgttc tcgtcagcaa aagagccgtt
     2101 catttcaata aaccgggcga cctcagccat cccttcctga ttttccgctt tccagcgttc
     2161 ggcacgcaga cgacgggctt cattctgcat ggttgtgctt accagaccgg agatattgac
     2221 atcatatatg ccttgagcaa ctgatagctg tcgctgtcaa ctgtcactgt aatacgctgc
     2281 ttcatagcat acctcttttt gacatacttc gggtatacat atcagtatat attcttatac
     2341 cgcaaaaatc agcgcgcaaa tacgcatact gttatctggc ttttagtaag ccggatccac
     2401 gcggcgttta cgccccccct gccactcatc gcagtactgt tgtaattcat taagcattct
     2461 gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac
     2521 cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat
     2581 attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa
     2641 catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc
     2701 ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga
     2761 aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac
     2821 cagctcaccg tctttcattg ccatacggaa ttccggatga gcattcatca ggcgggcaag
     2881 aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc
     2941 cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc
     3001 aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt
     3061 ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag
     3121 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa cgtctcattt
     3181 tcgccaaaag ttggcccagg gcttcccggt atcaacaggg acaccaggat ttatttattc
     3241 tgcgaagtga tcttccgtca caggtattta ttcggcgcaa agtgcgtcgg gtgatgctgc
     3301 caacttagtc gactacaggt cactaatacc atctaagtag ttgattcata gtgactggat
     3361 atgttgtgtt ttacagtatt atgtagtctg ttttttatgc aaaatctaat ttaatatatt
     3421 gatatttata tcattttacg tttctcgttc agctttcttg tacaaagtgg tgataaacgg
     3481 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     3541 ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg
     3601 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     3661 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     3721 caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta
     3781 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3841 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3901 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3961 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4021 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4081 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4141 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4201 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4261 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4321 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4381 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4441 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     4501 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4561 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     4621 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4681 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4741 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4801 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4861 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4921 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4981 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5041 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5101 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     5161 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5221 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5281 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5341 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5401 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc
     5461 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5521 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5581 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5641 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5701 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     5761 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5821 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5881 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5941 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     6001 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     6061 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     6121 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     6181 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6241 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6301 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6361 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6421 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6481 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6541 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6601 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6661 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6721 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6781 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6841 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6901 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6961 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     7021 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     7081 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     7141 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     7201 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     7261 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7321 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7381 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7441 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7501 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7561 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7621 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7681 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7741 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7801 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7861 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     7921 c
//
