LOCUS       40924_9796        6693 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Promoterless Gateway(R) destination vector for cloning of mammalian 
            promoters and genes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6693)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6693)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6693
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..114
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    144..268
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(700..1002)
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(1352..2005)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG"
     promoter        complement(2006..2108)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     protein_bind    complement(2286..2410)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2436..2477
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    2561..2609
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2811..3239
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3253..3582
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        3630..3677
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             3696..4091
                     /codon_start=1
                     /label=BSD
                     /note="blasticidin S deaminase"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4252..4385
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4422..4438)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4446..4462)
                     /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(4470..4500)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4515..4536)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4824..5412)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5586..6443)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6444..6548)
                     /label=AmpR promoter
ORIGIN
        1 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tctctggcta
       61 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc
      121 caagctggct agttaagcta tcaacaagtt tgtacaaaaa agctgaacga gaaacgtaaa
      181 atgatataaa tatcaatata ttaaattaga ttttgcataa aaaacagact acataatact
      241 gtaaaacaca acatatccag tcactatgaa tcaactactt agatggtatt agtgacctgt
      301 agtcgaccga cagccttcca aatgttcttc gggtgatgct gccaacttag tcgaccgaca
      361 gccttccaaa tgttcttctc aaacggaatc gtcgtatcca gcctactcgc tattgtcctc
      421 aatgccgtat taaatcataa aaagaaataa gaaaaagagg tgcgagcctc ttttttgtgt
      481 gacaaaataa aaacatctac ctattcatat acgctagtgt catagtcctg aaaatcatct
      541 gcatcaagaa caatttcaca actcttatac ttttctctta caagtcgttc ggcttcatct
      601 ggattttcag cctctatact tactaaacgt gataaagttt ctgtaatttc tactgtatcg
      661 acctgcagac tggctgtgta taagggagcc tgacatttat attccccaga acatcaggtt
      721 aatggcgttt ttgatgtcat tttcgcggtg gctgagatca gccacttctt ccccgataac
      781 ggagaccggc acactggcca tatcggtggt catcatgcgc cagctttcat ccccgatatg
      841 caccaccggg taaagttcac gggagacttt atctgacagc agacgtgcac tggccagggg
      901 gatcaccatc cgtcgcccgg gcgtgtcaat aatatcactc tgtacatcca caaacagacg
      961 ataacggctc tctcttttat aggtgtaaac cttaaactgc atttcaccag cccctgttct
     1021 cgtcagcaaa agagccgttc atttcaataa accgggcgac ctcagccatc ccttcctgat
     1081 tttccgcttt ccagcgttcg gcacgcagac gacgggcttc attctgcatg gttgtgctta
     1141 ccagaccgga gatattgaca tcatatatgc cttgagcaac tgatagctgt cgctgtcaac
     1201 tgtcactgta atacgctgct tcatagcata cctctttttg acatacttcg ggtatacata
     1261 tcagtatata ttcttatacc gcaaaaatca gcgcgcaaat acgcatactg ttatctggct
     1321 tttagtaagc cggatccacg cggcgtttac gccccccctg ccactcatcg cagtactgtt
     1381 gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa
     1441 tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg
     1501 gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg
     1561 gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt
     1621 caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt
     1681 attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt
     1741 gaacactatc ccatatcacc agctcaccgt ctttcattgc catacggaat tccggatgag
     1801 cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct
     1861 ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag
     1921 caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg
     1981 tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat ctcgataact
     2041 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
     2101 gccgatcaac gtctcatttt cgccaaaagt tggcccaggg cttcccggta tcaacaggga
     2161 caccaggatt tatttattct gcgaagtgat cttccgtcac aggtatttat tcggcgcaaa
     2221 gtgcgtcggg tgatgctgcc aacttagtcg actacaggtc actaatacca tctaagtagt
     2281 tgattcatag tgactggata tgttgtgttt tacagtatta tgtagtctgt tttttatgca
     2341 aaatctaatt taatatattg atatttatat cattttacgt ttctcgttca gctttcttgt
     2401 acaaagtggt tgatctagag ggcccgcggt tcgaaggtaa gcctatccct aaccctctcc
     2461 tcggtctcga ttctacgcgt accggttagt aatgagttta aacgggggag gctaactgaa
     2521 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     2581 aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     2641 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     2701 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     2761 aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg
     2821 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2881 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2941 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3001 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3061 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3121 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3181 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3241 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3301 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3361 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3421 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3481 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     3541 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     3601 atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg
     3661 gcatagtata atacgacaag gtgaggaact aaaccatggc caagcctttg tctcaagaag
     3721 aatccaccct cattgaaaga gcaacggcta caatcaacag catccccatc tctgaagact
     3781 acagcgtcgc cagcgcagct ctctctagcg acggccgcat cttcactggt gtcaatgtat
     3841 atcattttac tgggggacct tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg
     3901 cagctggcaa cctgacttgt atcgtcgcga tcggaaatga gaacaggggc atcttgagcc
     3961 cctgcggacg gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa gccatagtga
     4021 aggacagtga tggacagccg acggcagttg ggattcgtga attgctgccc tctggttatg
     4081 tgtgggaggg ctaagcactt cgtggccgag gagcaggact gacacgtgct acgagatttc
     4141 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     4201 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     4261 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     4321 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     4381 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     4441 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4501 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     4561 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4621 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4681 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4741 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4801 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4861 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4921 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4981 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5041 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5101 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5161 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5221 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5281 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5341 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5401 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5461 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5521 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5581 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5641 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5701 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5761 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5821 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5881 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5941 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6001 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6061 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6121 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6181 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6241 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6301 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6361 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6421 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6481 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6541 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg
     6601 atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     6661 tgctccctgc ttgtgtgttg gaggtcgctg agt
//
