LOCUS       Exported                6125 bp ds-DNA     circular SYN 08-JUL-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-Gateway
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6125)
  AUTHORS   White MD, Milne RV, Nolan MF
  TITLE     A Molecular Toolbox for Rapid Generation of Viral Vectors to Up- or 
            Down-Regulate Neuronal Gene Expression in vivo.
  JOURNAL   Front Mol Neurosci. 2011;4:8. Epub 2011 Jul 4.
  PUBMED    21772812
REFERENCE   2  (bases 1 to 6125)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 8, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6125
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   56..185
                     /label=AAV2 ITR
     polyA_signal    347..468
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(384..403)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     438..457
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    511..1099
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             617..628
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1026..1046
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    1119..1243
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1371..1390)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1916..2575)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2490..2509
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2629..2659)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(2684..2808)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2950..3227)
                     /label=chicken beta-actin promoter
     repeat_region   3538..3667
                     /label=AAV2 ITR
     rep_origin      3739..4194
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3826..3845)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4036..4057
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4220..4324
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4325..5185
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4543..4562)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5356..5944
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5845..5864
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6098..6115
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg
       61 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
      121 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
      181 ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa
      241 gatccttatc gattttacca catttgtaga ggttttactt gctttaaaaa acctcccaca
      301 tctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
      361 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
      421 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgctc
      481 gaagcggccg cccgggtcga cggtatcgat gcggggaggc ggcccaaagg gagatccgac
      541 tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag agccggcagc aggccgcggg
      601 aaggaaggtc cgctggattg agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa
      661 tccaggtggc aacacaggcg agcagccaag gaaaggacga tgatttcccc gacaacacca
      721 cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga
      781 tgagttccgc cgtggcaata gggaggggga aagcgaaagt cccggaaagg agctgacagg
      841 tggtggcaat gccccaacca gtgggggttg cgtcagcaaa cacagtgcac accacgccac
      901 gttgcctgac aacgggccac aactcctcat aaagagacag caaccaggat ttatacaagg
      961 aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt
     1021 atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt
     1081 tgtaatccag aggttgatta tcgataagct tgatatcaac cactttgtac aagaaagctg
     1141 aacgagaaac gtaaaatgat ataaatatca atatattaaa ttagattttg cataaaaaac
     1201 agactacata atactgtaaa acacaacata tccagtcact atggtcgacc tgcagactgg
     1261 ctgtgtataa gggagcctga catttatatt ccccagaaca tcaggttaat ggcgtttttg
     1321 tggctgagat cagccacttc ttccccgata acggagaccg gcacactggc catatcggtg
     1381 gtcatcatgc gccagctttc atccccgata tgcaccaccg ggtaaagttc acgggagact
     1441 ttatctgaca gcagacgtgc actggccagg gggatcacca tccgtcgccc gggcgtgtca
     1501 ataatatcac tctgtacatc cacaaacaga cgataacggc tctctctttt ataggtgtaa
     1561 accttaaact gcatttcacc agcccctgtt ctcgtcagca aaagagccgt tcatttcaat
     1621 aaaccgggcg acctcagcca tcccttcctg attttccgct ttccagcgtt cggcacgcag
     1681 acgacgggct tcattctgca tggttgtgct taccagaccg gagatattga catcatatat
     1741 gccttgagca actgatagct gtcgctgtca actgtcactg taatacgctg cttcatagca
     1801 tacctctttt tgacatactt cgggtataca tatcagtata tattcttata ccgcaaaaat
     1861 cagcgcgcaa atacgcatac tgttatctgg cttttagtaa gccggatcca cgcgtttacg
     1921 ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga
     1981 agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt
     2041 gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt
     2101 ttaaatcaaa actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa
     2161 taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata
     2221 tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag
     2281 tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt
     2341 ctttcattgc catacggaat tccggatgag cattcatcag gcgggcaaga atgtgaataa
     2401 aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca
     2461 gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt
     2521 tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag
     2581 cttccttagc tcctgaaaat ctcgacggat cctaactcaa aatccacaca ttatacgagc
     2641 cggaagcata aagtgtaaag cctggggtgc ctaatgcggc cgccatagtg actggatatg
     2701 ttgtgtttta cagtattatg tagtctgttt tttatgcaaa atctaattta atatattgat
     2761 atttatatca ttttacgttt ctcgttcagc ttttttgtac aaacttgttg atgctaatta
     2821 cagcccggag gagaagggcc gtcccgcccg ctcacctgtg ggagtaacgc ggtcagtcag
     2881 agccggggcg ggcggcgcga ggcggcggcg gagcggggca cggggcgaag gcagcgtcgc
     2941 agcgactccc gcccgccgcg cgcttcgctt tttatagggc cgccgccgcc gccgcctcgc
     3001 cataaaagga aactttcgga gcgcgccgct ctgattggct gccgccgcac ctctccgcct
     3061 cgccccgccc cgcccctcgc cccgccccgc cccgcctggc gcgcgccccc cccccccccc
     3121 cgcccccatc gctgcacaaa ataattaaaa aataaataaa tacaaaattg ggggtgggga
     3181 ggggggggag atggggagag tgaagcagaa cgtggggctc acctcgacca tggtaatagc
     3241 gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc
     3301 ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg
     3361 atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca
     3421 atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg tcaatggtac
     3481 tcttcctaga gcatggctac gtagataagt agcatggcgg gttaatcatt aactacaagg
     3541 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     3601 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     3661 cgcgcagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     3721 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3781 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3841 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3901 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3961 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4021 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4081 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     4141 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     4201 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4261 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4321 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4381 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4441 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4501 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4561 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4621 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     4681 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4741 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     4801 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     4861 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     4921 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     4981 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5041 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5101 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5161 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     5221 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5281 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5341 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5401 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5461 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5521 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5581 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5641 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5701 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5761 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5821 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5881 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5941 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6001 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     6061 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     6121 ggaag
//
