LOCUS       40924_9756        7519 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Gateway(R) destination vector for in vivo or in vitro detection of 
            mammalian gene expression using a C-terminal beta-lactamase 
            reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7519)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7519)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7519
                     /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             2630..3421
                     /codon_start=1
                     /label=bla(M)
                     /note="beta-lactamase lacking the signal sequence"
                     /translation="MDPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMM
                     STFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAIT
                     MSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMA
                     TTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGII
                     AALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     polyA_signal    3504..3552
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3754..4182
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4196..4525
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        4573..4620
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             4639..5034
                     /codon_start=1
                     /label=BSD
                     /note="blasticidin S deaminase"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5195..5328
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5365..5381)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5389..5405)
                     /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(5413..5443)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5458..5479)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5767..6352)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6526..7383)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7384..7488)
                     /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 ggatccgtcg 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 tttgtgatgg
     1741 cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg
     1801 ggcgtaaaga tctggatccg gcttactaaa agccagataa cagtatgcgt atttgcgcgc
     1861 tgatttttgc ggtataagaa tatatactga tatgtatacc cgaagtatgt caaaaagagg
     1921 tatgctatga agcagcgtat tacagtgaca gttgacagcg acagctatca gttgctcaag
     1981 gcatatatga tgtcaatatc tccggtctgg taagcacaac catgcagaat gaagcccgtc
     2041 gtctgcgtgc cgaacgctgg aaagcggaaa atcaggaagg gatggctgag gtcgcccggt
     2101 ttattgaaat gaacggctct tttgctgacg agaacagggg 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 ggctccctta tacacagcca gtctgcaggt cgaccatagt gactggatat gttgtgtttt
     2521 acagtattat gtagtctgtt ttttatgcaa aatctaattt aatatattga tatttatatc
     2581 attttacgtt tctcgttcag ctttcttgta caaagtggtt gatgctgtta tggacccaga
     2641 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     2701 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     2761 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     2821 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     2881 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     2941 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     3001 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     3061 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     3121 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     3181 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     3241 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     3301 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     3361 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     3421 gtaaccggtt agtaatgagt ttaaacgggg gaggctaact gaaacacgga aggagacaat
     3481 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg
     3541 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     3601 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     3661 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag
     3721 atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     3781 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     3841 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3901 taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3961 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     4021 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     4081 tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt
     4141 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     4201 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4261 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4321 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     4381 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     4441 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     4501 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     4561 tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac
     4621 aaggtgagga actaaaccat ggccaagcct ttgtctcaag aagaatccac cctcattgaa
     4681 agagcaacgg ctacaatcaa cagcatcccc atctctgaag actacagcgt cgccagcgca
     4741 gctctctcta gcgacggccg catcttcact ggtgtcaatg tatatcattt tactggggga
     4801 ccttgtgcag aactcgtggt gctgggcact gctgctgctg cggcagctgg caacctgact
     4861 tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg acggtgccga
     4921 caggtgcttc tcgatctgca tcctgggatc aaagccatag tgaaggacag tgatggacag
     4981 ccgacggcag ttgggattcg tgaattgctg ccctctggtt atgtgtggga gggctaagca
     5041 cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt
     5101 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5161 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     5221 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5281 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     5341 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5401 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5461 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5521 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5581 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5641 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5701 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5761 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5821 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5881 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5941 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6001 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6061 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6121 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6181 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6241 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6301 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6361 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6421 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6481 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6541 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6601 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6661 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6721 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6781 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6841 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6901 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6961 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7021 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7081 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7141 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7201 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7261 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7321 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7381 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7441 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7501 aaaagtgcca cctgacgtc
//
