LOCUS       40924_9781        7564 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Gateway(R) destination vector for in vivo or in vitro detection of 
            mammalian gene expression using an N-terminal beta-lactamase 
            reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7564)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7564)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7564
                     /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"
     CDS             911..1702
                     /codon_start=1
                     /label=bla(M)
                     /note="beta-lactamase lacking the signal sequence"
                     /translation="MDPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMM
                     STFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAIT
                     MSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMA
                     TTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGII
                     AALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     protein_bind    1712..1836
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1861..1891
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1945..2601
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2946..3248
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(3292..3416)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3424..3465
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    3549..3597
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3799..4227
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4241..4570
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        4618..4665
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             4684..5079
                     /codon_start=1
                     /label=BSD
                     /note="blasticidin S deaminase"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5240..5373
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5410..5426)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5434..5450)
                     /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(5458..5488)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5503..5524)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5812..6397)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6571..7428)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7429..7533)
                     /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 atggacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
      961 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     1021 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     1081 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     1141 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     1201 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     1261 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     1321 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     1381 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     1441 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     1501 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     1561 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     1621 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     1681 tgcctcactg attaagcatt ggctgttatc aacaagtttg tacaaaaaag ctgaacgaga
     1741 aacgtaaaat gatataaata tcaatatatt aaattagatt ttgcataaaa aacagactac
     1801 ataatactgt aaaacacaac atatccagtc actatggcgg ccgcattagg caccccaggc
     1861 tttacacttt atgcttccgg ctcgtataat gtgtggattt tgagttagga tccgtcgaga
     1921 ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat
     1981 atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc
     2041 tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag
     2101 cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa
     2161 ttccgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac
     2221 accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggagtgaata ccacgacgat
     2281 ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc
     2341 tatttcccta aagggtttat tgagaatatg tttttcgtct cagccaatcc ctgggtgagt
     2401 ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc cgttttcacc
     2461 atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat tcaggttcat
     2521 catgccgttt gtgatggctt ccatgtcggc agaatgctta atgaattaca acagtactgc
     2581 gatgagtggc agggcggggc gtaaagatct ggatccggct tactaaaagc cagataacag
     2641 tatgcgtatt tgcgcgctga tttttgcggt ataagaatat atactgatat gtatacccga
     2701 agtatgtcaa aaagaggtat gctatgaagc agcgtattac agtgacagtt gacagcgaca
     2761 gctatcagtt gctcaaggca tatatgatgt caatatctcc ggtctggtaa gcacaaccat
     2821 gcagaatgaa gcccgtcgtc tgcgtgccga acgctggaaa gcggaaaatc aggaagggat
     2881 ggctgaggtc gcccggttta ttgaaatgaa cggctctttt gctgacgaga acaggggctg
     2941 gtgaaatgca gtttaaggtt tacacctata aaagagagag ccgttatcgt ctgtttgtgg
     3001 atgtacagag tgatattatt gacacgcccg ggcgacggat ggtgatcccc ctggccagtg
     3061 cacgtctgct gtcagataaa gtctcccgtg aactttaccc ggtggtgcat atcggggatg
     3121 aaagctggcg catgatgacc accgatatgg ccagtgtgcc ggtctccgtt atcggggaag
     3181 aagtggctga tctcagccac cgcgaaaatg acatcaaaaa cgccattaac ctgatgttct
     3241 ggggaatata aatgtcaggc tcccttatac acagccagtc tgcaggtcga ccatagtgac
     3301 tggatatgtt gtgttttaca gtattatgta gtctgttttt tatgcaaaat ctaatttaat
     3361 atattgatat ttatatcatt ttacgtttct cgttcagctt tcttgtacaa agtggttgat
     3421 aacgggaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggttagtaa
     3481 tgagtttaaa cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     3541 cgctatgacg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata
     3601 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     3661 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     3721 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg
     3781 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3841 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3901 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc
     3961 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     4021 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4081 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4141 tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc
     4201 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     4261 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4321 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4381 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4441 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     4501 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4561 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt
     4621 gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa
     4681 accatggcca agcctttgtc tcaagaagaa tccaccctca ttgaaagagc aacggctaca
     4741 atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct ctctagcgac
     4801 ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg tgcagaactc
     4861 gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc
     4921 ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt gcttctcgat
     4981 ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac ggcagttggg
     5041 attcgtgaat tgctgccctc tggttatgtg tgggagggct aagcacttcg tggccgagga
     5101 gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5161 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5221 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     5281 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5341 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     5401 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5461 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5521 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5581 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5641 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5701 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5761 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5821 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5881 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5941 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6001 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6061 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6121 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6181 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6241 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6301 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg
     6361 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6421 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6481 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6541 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6601 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6661 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6721 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6781 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6841 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6901 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6961 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7021 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7081 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7141 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7201 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7261 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7321 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7381 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7441 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7501 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7561 cgtc
//
