LOCUS       40924_25296        5038 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Gateway(R) destination vector for stable constitutive expression of 
            C-terminally V5-6xHis-tagged proteins in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5038)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5038)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5038
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..548
                     /label=OpIE-2 promoter
                     /note="strong constitutive baculovirus promoter for insect
                     cell expression"
     protein_bind    602..726
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        751..781
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             835..1491
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             1836..2138
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2182..2306)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2341..2382
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     CDS             2392..2409
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     polyA_signal    2427..2556
                     /label=OpIE-2 poly(A) signal
                     /note="baculovirus polyadenylation signal"
     rep_origin      complement(2684..3272)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        3364..3463
                     /label=gp64 promoter
                     /note="promoter for the baculovirus gp64 glycoprotein gene"
     promoter        3499..3546
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             3565..3960
                     /codon_start=1
                     /label=BSD
                     /note="blasticidin S deaminase"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVAGAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             4083..4940
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 catgatgata aacaatgtat ggtgctaatg ttgcttcaac aacaattctg ttgaactgtg
       61 ttttcatgtt tgccaacaag cacctttata ctcggtggcc tccccaccac caactttttt
      121 gcactgcaaa aaaacacgct tttgcacgcg ggcccataca tagtacaaac tctacgtttc
      181 gtagactatt ttacataaat agtctacacc gttgtatacg ctccaaatac actaccacac
      241 attgaacctt tttgcagtgc aaaaaagtac gtgtcggcag tcacgtaggc cggccttatc
      301 gggtcgcgtc ctgtcacgta cgaatcacat tatcggaccg gacgagtgtt gtcttatcgt
      361 gacaggacgc cagcttcctg tgttgctaac cgcagccgga cgcaactcct tatcggaaca
      421 ggacgcgcct ccatatcagc cgcgcgttat ctcatgcacg tgaccggaca cgaggcgccc
      481 gtcccgctta tcgcgcctat aaatacagcc cgcaacgatc tggtaaacac agttgaacag
      541 catctgttcg aatttaaagc ttgatatcga attcctgcag cccagcgctg gatcctcgat
      601 cacaagtttg tacaaaaaag ctgaacgaga aacgtaaaat gatataaata tcaatatatt
      661 aaattagatt ttgcataaaa aacagactac ataatactgt aaaacacaac atatccagtc
      721 actatggcgg ccgcattagg caccccaggc tttacacttt atgcttccgg ctcgtataat
      781 gtgtggattt tgagttagga tccgtcgaga ttttcaggag ctaaggaagc taaaatggag
      841 aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
      901 gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg
      961 gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt
     1021 cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga cggtgagctg
     1081 gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt
     1141 tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa
     1201 gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg
     1261 tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat
     1321 atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag
     1381 gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc
     1441 agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc gtaaacgcgt
     1501 ggatccggct tactaaaagc cagataacag tatgcgtatt tgcgcgctga tttttgcggt
     1561 ataagaatat atactgatat gtatacccga agtatgtcaa aaagaggtat gctatgaagc
     1621 agcgtattac agtgacagtt gacagcgaca gctatcagtt gctcaaggca tatatgatgt
     1681 caatatctcc ggtctggtaa gcacaaccat gcagaatgaa gcccgtcgtc tgcgtgccga
     1741 acgctggaaa gcggaaaatc aggaagggat ggctgaggtc gcccggttta ttgaaatgaa
     1801 cggctctttt gctgacgaga acaggggctg gtgaaatgca gtttaaggtt tacacctata
     1861 aaagagagag ccgttatcgt ctgtttgtgg atgtacagag tgatattatt gacacgcccg
     1921 ggcgacggat ggtgatcccc ctggccagtg cacgtctgct gtcagataaa gtctcccgtg
     1981 aactttaccc ggtggtgcat atcggggatg aaagctggcg catgatgacc accgatatgg
     2041 ccagtgtgcc ggtctccgtt atcggggaag aagtggctga tctcagccac cgcgaaaatg
     2101 acatcaaaaa cgccattaac ctgatgttct ggggaatata aatgtcaggc tcccttatac
     2161 acagccagtc tgcaggtcga ccatagtgac tggatatgtt gtgttttaca gtattatgta
     2221 gtctgttttt tatgcaaaat ctaatttaat atattgatat ttatatcatt ttacgtttct
     2281 cgttcagctt tcttgtacaa agtggtgatc gacccgggtc tagagggccc gcggttcgaa
     2341 ggtaagccta tccctaaccc tctcctcggt ctcgattcta cgcgtaccgg tcatcatcac
     2401 catcaccatt gagtttatct gactaaatct tagtttgtat tgtcatgttt taatacaata
     2461 tgttatgttt aaatatgttt ttaataaatt ttataaaata atttcaactt ttattgtaac
     2521 aacattgtcc atttacacac tcctttcaag cgcgtgggat cgatgctcac tcaaaggcgg
     2581 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     2641 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     2701 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2761 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2821 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     2881 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     2941 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3001 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3061 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3121 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3181 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3241 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3301 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgccc ttgttccgaa
     3361 gggttgtgtc acgtaggcca gataacggtc gggtatataa gatgcctcaa tgctactagt
     3421 aaatcagtca caccaaggct tcaataagga acacacaagc aagccctttg agtcaagggc
     3481 tgccgggctg cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     3541 tacgacaagg tgaggaacta aaccatggcc aagcctttgt ctcaagaaga atccaccctc
     3601 attgaaagag caacggctac aatcaacagc atccccatct ctgaagacta cagcgtcgcc
     3661 ggcgcagctc tctctagcga cggccgcatc ttcactggtg tcaatgtata tcattttact
     3721 gggggacctt gcgcagaact cgtggtgctg ggcactgctg ctgctgcggc agctggcaac
     3781 ctgacttgta tcgtcgcgat cggaaatgag aacaggggca tcttgagccc ctgcggacgg
     3841 tgccgacagg ttcttctcga tctgcatcct gggatcaaag ccatagtgaa ggacagtgat
     3901 ggacagccga cggcagttgg gattcgtgaa ttgctgccct ctggttatgt gtgggagggc
     3961 taagcacttc gtggccgagg agcaggactg acacgtcccg ggagatctgc atgtctacta
     4021 aactcacaaa ttagagcttc aatttaatta tatcagttat tacccattga aaaaggaaga
     4081 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4141 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4201 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4261 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4321 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4381 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4441 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4501 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4561 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4621 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     4681 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     4741 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     4801 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     4861 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4921 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     4981 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatct
//
