LOCUS       40924_9761        6809 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Mammalian Gateway(R) destination vector for C-terminal tagging of a 
            protein with the Lumio(TM) tetracysteine tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6809)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6809)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6809
                     /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    915..1039
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1064..1094
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1148..1804
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2149..2451
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2495..2619)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2633..2674
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     CDS             2693..2710
                     /codon_start=1
                     /label=tetracysteine tag
                     /note="tetracysteine peptide that binds biarsenical
                     labeling reagents"
                     /translation="CCPGCC"
     polyA_signal    2794..2842
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3044..3472
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3486..3815
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        3863..3910
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             3929..4324
                     /codon_start=1
                     /label=BSD
                     /note="blasticidin S deaminase"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4485..4618
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4655..4671)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4679..4695)
                     /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(4703..4733)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4748..4769)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5057..5642)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5816..6673)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6674..6778)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 taagctgagc atcaacaagt ttgtacaaaa aagctgaacg agaaacgtaa aatgatataa
      961 atatcaatat attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac
     1021 aacatatcca gtcactatgg cggccgcatt aggcacccca ggctttacac tttatgcttc
     1081 cggctcgtat aatgtgtgga ttttgagtta ggatccggcg agattttcag gagctaagga
     1141 agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg
     1201 taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca
     1261 gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc
     1321 ctttattcac attcttgccc gcctgatgaa tgctcatccg gaattccgta tggcaatgaa
     1381 agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca
     1441 aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca
     1501 catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt
     1561 tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt
     1621 aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac
     1681 gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg tctgtgatgg
     1741 cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg
     1801 ggcgtaaaga tctggatccg gcttactaaa agccagataa cagtatgcgt atttgcgcgc
     1861 tgatttttgc ggtataagaa tatatactga tatgtatacc cgaagtatgt caaaaagagg
     1921 tgtgctatga agcagcgtat tacagtgaca gttgacagcg acagctatca gttgctcaag
     1981 gcatatatga tgtcaatatc tccggtctgg taagcacaac catgcagaat gaagcccgtc
     2041 gtctgcgtgc cgaacgctgg aaagcggaaa atcaggaagg gatggctgag gtcgcccggt
     2101 ttattgaaat gaacggctct tttgctgacg agaacaggga ctggtgaaat gcagtttaag
     2161 gtttacacct ataaaagaga gagccgttat cgtctgtttg tggatgtaca gagtgatatt
     2221 attgacacgc ccgggcgacg gatggtgatc cccctggcca gtgcacgtct gctgtcagat
     2281 aaagtctccc gtgaacttta cccggtggtg catatcgggg atgaaagctg gcgcatgatg
     2341 accaccgata tggccagtgt gccggtctcc gttatcgggg aagaagtggc tgatctcagc
     2401 caccgcgaaa atgacatcaa aaacgccatt aacctgatgt tctggggaat ataaatgtca
     2461 ggctccgtta tacacagcca gtctgcaggt cgaccatagt gactggatat gttgtgtttt
     2521 acagtattat gtagtctgtt ttttatgcaa aatctaattt aatatattga tatttatatc
     2581 attttacgtt tctcgttcag ctttcttgta caaagtggtt gatgctgtta acgggaagcc
     2641 tatccctaac cctctcctcg gtctcgattc tacgcgtacc ggtgctggtg gctgttgtcc
     2701 tggctgttgc ggtggcggct agtaatgagt ttaaacgggg gaggctaact gaaacacgga
     2761 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     2821 acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     2881 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     2941 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     3001 gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     3061 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3121 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3181 cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc
     3241 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3301 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3361 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt
     3421 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3481 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3541 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3601 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3661 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3721 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     3781 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     3841 ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt
     3901 ataatacgac aaggtgagga actaaaccat ggccaagcct ttgtctcaag aagaatccac
     3961 cctcattgaa agagcaacgg ctacaatcaa cagcatcccc atctctgaag actacagcgt
     4021 cgccagcgca gctctctcta gcgacggccg catcttcact ggtgtcaatg tatatcattt
     4081 tactggggga ccttgtgcag aactcgtggt gctgggcact gctgctgctg cggcagctgg
     4141 caacctgact tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg
     4201 acggtgccga caggtgcttc tcgatctgca tcctgggatc aaagccatag tgaaggacag
     4261 tgatggacag ccgacggcag ttgggattcg tgaattgctg ccctctggtt atgtgtggga
     4321 gggctaagca cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca
     4381 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     4441 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     4501 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4561 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     4621 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     4681 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4741 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     4801 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4861 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4921 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4981 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5041 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5101 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5161 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5221 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5281 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5341 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5401 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5461 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     5521 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5581 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5641 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5701 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5761 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5821 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5881 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5941 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6001 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6061 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6121 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6181 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6241 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6301 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6361 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6421 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6481 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6541 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6601 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6661 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6721 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6781 catttccccg aaaagtgcca cctgacgtc
//
