LOCUS Exported 8386 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pAAV-hRREB1-RFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8386) AUTHORS Nathanson JL, Jappelli R, Scheeff ED, Manning G, Obata K, Brenner S, Callaway EM TITLE Short Promoters in Viral Vectors Drive Selective Expression in Mammalian Inhibitory Neurons, but do not Restrict Activity to Specific Inhibitory Cell-Types. JOURNAL Front Neural Circuits. 2009 . 3():19. PUBMED 19949461 REFERENCE 2 (bases 1 to 8386) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8386 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 7..136 /label=AAV2 ITR promoter 2963..3001 /label=minimal CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" intron 3060..3535 /label=-beta-globin intron /note="internally truncated intron from human beta-globin" regulatory 3611..3620 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3617..4294 /codon_start=1 /product="rapidly maturing tetrameric variant of DsRed fluorescent protein (Bevis and Glick, 2002)" /label=DsRed-Express /note="mammalian codon-optimized" /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH LFL" primer_bind complement(3747..3765) /label=mCherry-R /note="mCherry, reverse primer" primer_bind complement(3797..3817) /label=DsRed1-N /note="DsRed1, reverse primer" primer_bind 4015..4034 /label=mCherry-F /note="mCherry, forward primer" primer_bind 4209..4232 /label=DsRed1-C /note="DsRed1, forward primer" promoter complement(4324..4342) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(4325..4342) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(4362..4379) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4368..4479 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 5170..5683 /direction=RIGHT /label=M13 ori /note="M13 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5257..5276) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5467..5488 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(6164..6183) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 6283..6305 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(6343..6361) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 6429..6533 /gene="bla" /label=AmpR promoter CDS 6534..7394 /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(6752..6771) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7565..8153 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8054..8073 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 8307..8324 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 cagcagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg 61 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc 121 atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc tacgtagcca 181 tgctctagag gatccttcga aaagcttaca atggcttaca cctgtaatcc cagcaatttg 241 ggaggccaag gtgggagaat cacttgagcc ctagagcctg agaccagcct gggcaacata 301 gcaagacctc ctctctacta aaacttttaa aattaactag atgtggtgac atgcactgta 361 gtcccaccta cttgggaggc taaaggtgga ggatcacttg agaccaggaa ttcaaagctg 421 cagtgagttg tgatcccacc actgaactcc agcccgggca acagagtgat gctctgtctc 481 caaaaaaaaa aaaaaattaa aaaaagttgt agtaaaactg ttttcaatct tttttgttgc 541 cagtattttc tagatggcat tatacaggtg gcaatttgtc atgaaaattc atatagcatt 601 ggagatttgc taaattatgc ataaaattcc aattttgata tttttcctat tcaaaactta 661 tccctgaatg gggagaccag gaaaggctag ctcctgaatc agagacaaag tgagtaaata 721 aagtagcatt ccccacaggg cccttcctct gccctcattg gaacacatta caacttagaa 781 acacaagaca cctattctat gatggcaatg tgaactatta ttcctgtcta gagacctgcc 841 catctcctcc acgactggcc tgtcttcaaa aaagcatttg ataatacatt tgggggcggc 901 ggggttgggg gaggctcaga aaaatgcaac cgctaaaatt tctctggatg gagttttctt 961 cagtaaacag tagaccctgg gaaacataac aggttccgaa aacttaattt tgatgactga 1021 aagtcagaga aacagccagc tgaatcagct ccttcttggg cagtggttta gcctacttat 1081 ctcatatcag acctaagggt ccagctgctt ttccttggtt ctaaagagtt ctattctggc 1141 caggacccat cccgagacag acacttctgt ctgcagccct gaaaactaga ggtcaatgga 1201 agggctcaaa ttaaaggccc acagttgggc ttacaactct ccatggtgtt caagccttct 1261 cagaggagct ggtgtggtgg agccagggtc aaaggtcata gtaaatagaa agtagccagc 1321 tttctgcctg cataacatca tcttgcctaa ccagactcaa acgcttaatt ctactgtagt 1381 attctttgga cattcttctt tagaactacg ggggcggggg ggggggggcg tgtttatttt 1441 caacaaccac aatcatcctt atctttctga agatgtttcc aaaggctgag aacagacccc 1501 tttcagaacc tcggtaggag aagtattaac agttggaaac atgtcagctg ctttatggag 1561 gaagcacagc ccaggcctgg tttctctggg ctccgtgata acttggggca aggcagccag 1621 tgcgaagcgt ggtggcagga gctggagaga tggagagcgg ctcggagaag ggcccctgct 1681 tttcagagtt gaaacaggag ccctctgaga gaaagagata ctgcgtgtgg cttctctgaa 1741 gccttggagg gcgacgtgga tgacacagca tcagccattc catcgcgggg tggggacaca 1801 gtggcggcag gaaccgtcct cagcccctgg cggccacggc agttccccgc tgaagcatcc 1861 tttgtggggt gtgggtaagg gagggggacc aaagtcgggt cggagttgag tttgttagac 1921 ggcgagcaca ctcagggcgg cagcccgagt aagaaggatg gccccgggag agaaacggct 1981 cgaagctagg ggcggggtga agcgcctgct ccttaatctc ctgccccatc atccactcgc 2041 cccacctgga tgcagagcga ggaccgcgtg gaaaggaaac ggcgcgggcg gggctccccc 2101 tcgcagatgc ctcggagacc cagaggggcg gcgtcaggcc agggccccga gccctacgcc 2161 ccccatcccc ccaactccag cgcctctgag gtgcccctct cgcaattccg aggcatcacc 2221 gtgtctcccc cgcctccgtt aaaatagtga aactgcggag tgaagtgatg gaaagccggg 2281 ggcggggagt cgcccgcggc gaggggctgc aggggagggg gcgcggtggc cgaataaggt 2341 ggcccgcgag caacgggacg attgggatcg cagccagtcg cggcctcccg gacccttccc 2401 ctcgggcagc ccggccctcc catggcccgg ggcgccttgg gagggcgtag ggcaaacctg 2461 gggctgcggt ggggggtggg gaggggactc cacctccatc catcccggga tggcaactgc 2521 ggtcaccctg ctaaagtcgg ggcgggggcg gcggtcctcc ccctcacccc ccccagtccg 2581 agcgccgccg ccgccgccgc cgccgccgcc gcggccgctc agtaacacgt ccccaggaga 2641 ctcgcaggag caacacgtga tgtgtctact tatcagggtg agagggggag agcgagactc 2701 cgagcgtgcg ggcgaggaag ctgggcgggg ggccgagacg ggggccggga cgacggctgg 2761 ggcgccgggg tcgccagctc ctcgcccacc tccctccctg tgggagaaga ggaggagggg 2821 aagtgactgc ggagttgatg aactttgcac atccagaaac gccattttga ttccttttcc 2881 ggaacaagtt tgcatctctc ctcggtctct ccctgaagcc caccggtccc caacgcatca 2941 tgccagggag ccaattaatt aaggtaggcg tgtacggtgg gaggtctata taagcagagc 3001 tggtttagtg aaccgtcaga tccgctagcg cgccgagctc ggatcctgag aacttcaggg 3061 tgagtctatg ggacccttga tgttttcttt ccccttcttt tctatggtta agttcatgtc 3121 ataggaaggg gagaagtaac agggtacaca tattgaccaa atcagggtaa ttttgcattt 3181 gtaattttaa aaaatgcttt cttcttttaa tatacttttt tgtttatctt atttctaata 3241 ctttccctaa tctctttctt tcagggcaat aatgatacaa tgtatcatgc ctctttgcac 3301 cattctaaag aataacagtg ataatttctg ggttaaggca atagcaatat ttctgcatat 3361 aaatatttct gcatataaat tgtaactgat gtaagaggtt tcatattgct aatagcagct 3421 acaatccagc taccattctg cttttatttt atggttggga taaggctgga ttattctgag 3481 tccaagctag gcccttttgc taatcatgtt catacctctt atcttcctcc cacagctcct 3541 gggcaacgtg ctggtctgtg tgctggccca tcactttggc aaagcacgtg ggccgcctcg 3601 gccaccggtc gccaccatgg cctcctccga ggacgtcatc aaggagttca tgcgcttcaa 3661 ggtgcgcatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg 3721 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc 3781 cttcgcctgg gacatcctgt ccccccagtt ccagtacggc tccaaggtgt acgtgaagca 3841 ccccgccgac atccccgact acaagaagct gtccttcccc gagggcttca agtgggagcg 3901 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga 3961 cggctccttc atctacaagg tgaagttcat cggcgtgaac ttcccctccg acggccccgt 4021 aatgcagaag aagactatgg gctgggaggc ctccaccgag cgcctgtacc cccgcgacgg 4081 cgtgctgaag ggcgagatcc acaaggccct gaagctgaag gacggcggcc actacctggt 4141 ggagttcaag tccatctaca tggccaagaa gcccgtgcag ctgcccggct actactacgt 4201 ggactccaag ctggacatca cctcccacaa cgaggactac accatcgtgg agcagtacga 4261 gcgcgccgag ggccgccacc acctgttcct gtagagttta aaccgtacgg gaaacggccc 4321 tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg tgccttctag 4381 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac 4441 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca 4501 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag 4561 cgcggccgct ctagagcatg gctacgtaga taagtagcat ggcgggttaa tcattaacta 4621 caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga 4681 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga 4741 gcgagcgcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt 4801 tgcgcagcct gaatggcgaa tggaattcca gacgattgag cgtcaaaatg taggtatttc 4861 catgagcgtt tttcctgttg caatggctgg cggtaatatt gttctggata ttaccagcaa 4921 ggccgatagt ttgagttctt ctactcaggc aagtgatgtt attactaatc aaagaagtat 4981 tgcgacaacg gttaatttgc gtgatggaca gactctttta ctcggtggcc tcactgatta 5041 taaaaacact tctcaggatt ctggcgtacc gttcctgtct aaaatccctt taatcggcct 5101 cctgtttagc tcccgctctg attctaacga ggaaagcacg ttatacgtgc tcgtcaaagc 5161 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca 5221 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct 5281 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt 5341 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac 5401 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct 5461 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt 5521 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac 5581 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aatttaaata tttgcttata 5641 caatcttcct gtttttgggg cttttctgat tatcaaccgg ggtacatatg attgacatgc 5701 tagttttacg attaccgttc atcgattctc ttgtttgctc cagactctca ggcaatgacc 5761 tgatagcctt tgtagagacc tctcaaaaat agctaccctc tccggcatga atttatcagc 5821 tagaacggtt gaatatcata ttgatggtga tttgactgtc tccggccttt ctcacccgtt 5881 tgaatcttta cctacacatt actcaggcat tgcatttaaa atatatgagg gttctaaaaa 5941 tttttatcct tgcgttgaaa taaaggcttc tcccgcaaaa gtattacagg gtcataatgt 6001 ttttggtaca accgatttag ctttatgctc tgaggcttta ttgcttaatt ttgctaattc 6061 tttgccttgc ctgtatgatt tattggatgt tggaattcct gatgcggtat tttctcctta 6121 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg 6181 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt 6241 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc 6301 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat 6361 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg 6421 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc 6481 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta 6541 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg 6601 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg 6661 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac 6721 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg 6781 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt 6841 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg 6901 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac 6961 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt 7021 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag 7081 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc 7141 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc 7201 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta 7261 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg 7321 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga 7381 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac 7441 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa 7501 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat 7561 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc 7621 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg 7681 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc 7741 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg 7801 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg 7861 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa 7921 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg 7981 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga 8041 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct 8101 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca 8161 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc 8221 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg 8281 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc 8341 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatg //