LOCUS Exported 7478 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION INTRSECT 2.0. ACCESSION . VERSION . KEYWORDS pAAV-Ef1a-Con/Fon oG SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7478) AUTHORS Hafner G, Witte M, Guy J, Subhashini N, Fenno LE, Ramakrishnan C, Kim YS, Deisseroth K, Callaway EM, Oberhuber M, Conzelmann KK, Staiger JF TITLE Mapping Brain-Wide Afferent Inputs of Parvalbumin-Expressing GABAergic Neurons in Barrel Cortex Reveals Local and Long-Range Circuit Motifs. JOURNAL Cell Rep. 2019 Sep 24;28(13):3450-3461.e8. doi: 10.1016/j.celrep.2019.08.064. PUBMED 31553913 REFERENCE 2 (bases 1 to 7478) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7478 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" promoter 231..1409 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 462..1400 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1357..1377 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" protein_bind 1416..1449 /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind 1500..1533 /label=loxN /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GTATACCT). loxN sites are compatible with each other, but incompatible with loxP or lox2272 sites." regulatory 2194..2203 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind complement(2208..2241) /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind complement(2292..2325) /label=loxN /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GTATACCT). loxN sites are compatible with each other, but incompatible with loxP or lox2272 sites." misc_feature 3605..4193 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3658..3678) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4076..4087) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 4225..4701 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4424..4443) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4741..4881 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4741..4870 /label=AAV2 ITR rep_origin 4956..5411 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5043..5062) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5253..5274 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5428..5447) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5547..5569 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5607..5625) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5693..5797 /gene="bla" /label=AmpR promoter CDS 5798..6658 /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(6016..6035) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6829..7417 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7318..7337 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt 61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 121 aggggttcct gcggccgcac gcgtaagctt tgcaaagatg gataaagttt taaacagaga 181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg 241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg 301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt 361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg 421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt 481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct 541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga 601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct 661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt 721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc 781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg 841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac 901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc 961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag 1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct 1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg 1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga 1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc 1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg 1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa 1381 gtttttttct tccatttcag gtgtcgtgag gatccataac ttcgtatagg atactttata 1441 cgaagttatg tagctggagc aatttgaaaa gattgttgta ttacctctag ctgcgcttaa 1501 taacttcgta tagtatacct tatacgaagt tatgtcgacc ctcctgtact cacctgctgt 1561 tggtgaagat gtcgcaggac atccccagcc ttggattctc aggcatccag attgtataat 1621 catggttggt ggaacagtaa gtggagctga cggccactcc ggagcagttt ccgccgggaa 1681 acactggact gtggagagat cggtcgtagg ggtccaggtc ggcgacagat ggggaaatga 1741 tgaccaggct ctcttttgtg gtcttcacag tgcgcagcca atgatagtct gggtaaggat 1801 tgtgcaggct ttcctcatag cgggggtcgc cagccatctt ccagttgtaa gctgcccgac 1861 aggcgtctgg tgttggccga aagtgttttc tcttgaaagt tgtggtgaca tatcccacaa 1921 aatttgtgta ggtttctgcc tcggtgacca caccagtgca tgtgaacccg ttcatcttaa 1981 tagcagagat atagcccact ttcagctcca tgtaggagaa tccgctcagg ttggtacagc 2041 cttcgtcctc gaccacgaga ttgttagggc atgacagatg gtgaatatcg ataggactcc 2101 aggggcccag cttgtcgggg atagtgtaga tagggaattt accaaagcag agtgggaaga 2161 cgagcagagg gacgaagagg agagcctgtg ggaccatggt ggctagcata acttcgtata 2221 aagtatccta tacgaagtta tttgccttaa cccagaaatt atcactgtta ttctttagat 2281 aggtgcaaag aataacttcg tataaggtat actatacgaa gttatggtac cgaagttcct 2341 attcttcaaa tagtatagga acttcaattg ttattggtgc aaagaatcac ttccttaagt 2401 ctttatactg cccaggaagt tcctattctt caaaaggtat aggaacttcg gcgcgcctta 2461 gagccgtgtc tcgcccccag atttatgact ttcccaactg ctgatgattt taccgctctg 2521 gggtgtgaca gacacttctc taccggtccc cctcagattg tgctgagttg gctcggagcg 2581 gttcactcgt cgccagcagg tcatcaggaa aatgatcagc atgagagcag tcagtgctcc 2641 agcggagagc agcacgtact tgccccaatt aggcagtccg agatcgacgc cactgatccg 2701 ttcgtgcacg tcgggcaggt ggacctccac aaaatcttcg gcctcgtcgc cgtttttgaa 2761 cactgtgctt gggtctgcga gaggatgcat caggggaatg acgcttgaca cgagcagttc 2821 catgtgctgc tggagcagac tagactgcat ctcggggatc agcacgttac catctggccc 2881 cagaatgata ccattaaaga agaccccgtt cacatgtggg tgacatcgac ccccgactcg 2941 caggcatccc ttactaggaa tgatttcatt ccaggttctc acagatttat aatgggcgtc 3001 tgcctccatc agtgtcttgt taaagatggt gtatgctttt ccgaagccag ggacgagctt 3061 cctcaggtgg ctcagcctcc tgaagctcac gctcttggta gtcataattg actccagagc 3121 atcgagacat tcctcccttt tcttcacgag ttcctcgacc accagatgtt cgatctcatc 3181 gcttcggaag tcgtgcagat tcacgagctg cccagggggg caccacttag tctcgttaga 3241 tgtctgcata gcgacccagg ttccgtccat cagccggagt cccagcacgc cacagagttt 3301 cagcttgcag gcacctttga ggctcttata cagccctctt tcatccacaa atccacaagt 3361 ctcgctgcct tttgaggccc tcttgcccct gtgggacgac atgacttaac cagttaacca 3421 tgttctctga acatgagaag ggctggccac tctccatcta gagaagttcc tatactattt 3481 gaagaatagg aacttcttgc cttaacccag aaattatcac tgttattctt tagaatagtg 3541 caaagagaag ttcctatacc ttttgaagaa taggaacttc gaattcgata tcaagcttat 3601 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 3661 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 3721 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 3781 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 3841 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 3901 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 3961 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct 4021 gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 4081 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 4141 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata 4201 ccgagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc agtgcctctc 4261 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc 4321 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg 4381 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc 4441 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt 4501 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa 4561 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc 4621 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc 4681 actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc gagcggccgc 4741 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 4801 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc 4861 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg 4921 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag 4981 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc 5041 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc 5101 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa 5161 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg 5221 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac 5281 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta 5341 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac 5401 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca 5461 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc 5521 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc 5581 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt 5641 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac 5701 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc 5761 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt 5821 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct 5881 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga 5941 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag 6001 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca 6061 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga 6121 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag 6181 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc 6241 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa 6301 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt 6361 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg 6421 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt 6481 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg 6541 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat 6601 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact 6661 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa 6721 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt 6781 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt 6841 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg 6901 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca 6961 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt 7021 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga 7081 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc 7141 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact 7201 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga 7261 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg 7321 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt 7381 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt 7441 acggttcctg gccttttgct ggccttttgc tcacatgt //