LOCUS Exported 7560 bp ds-DNA circular SYN 18-JUN-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS AAV-Flex-rev-oChIEF-tdTomato SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7560) AUTHORS Bell KA, Shim H, Chen CK, McQuiston AR TITLE Nicotinic excitatory postsynaptic potentials in hippocampal CA1 interneurons are predominantly mediated by nicotinic receptors that contain alpha4 and beta2 subunits. JOURNAL Neuropharmacology. 2011 Aug 25. PUBMED 21878344 REFERENCE 2 (bases 1 to 7560) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 18, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7560 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 55..74 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" protein_bind complement(109..142) /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(205..238) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS complement(256..1686) /codon_start=1 /product="tandem dimeric (pseudo-monomeric) derivative of DsRed (Shaner et al., 2004)" /label=tdTomato /note="mammalian codon-optimized" /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE RSEGRHHLFLYGMDELYK" protein_bind 2796..2829 /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 2900..2933 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 2975..3563 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3028..3048) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3446..3457) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3606..3727 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3617..3636) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3671..3690 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 3889..4018 /label=AAV2 ITR primer_bind complement(4084..4101) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4255..4843) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4335..4354) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5014..5874) /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 5637..5656 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5875..5979) /gene="bla" /label=AmpR promoter rep_origin complement(6005..6460) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6142..6163) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 6354..6373 /label=F1ori-R /note="F1 origin, reverse primer" repeat_region 6532..6661 /label=AAV2 ITR promoter 6974..7251 /label=chicken beta-actin promoter primer_bind 7531..7553 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" ORIGIN 1 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg 61 tgctggttat tgtgctgtct catcattttg gcaaagaatt ggatccccat aacttcgtat 121 aaagtatcct atacgaagtt atatcaaaat aggaagacca atgcttcacc atcgacccga 181 attgccaagc atcaccatcg acccataact tcgtataatg tatgctatac gaagttatac 241 taggatcccc tcgagctact tgtacagctc gtccatgccg tacaggaaca ggtggtggcg 301 gccctcggag cgctcgtact gttccacgat ggtgtagtcc tcgttgtggg aggtgatgtc 361 cagcttggtg tccacgtagt agtagccggg cagttgcacg ggcttcttgg ccatgtagat 421 ggtcttgaac tccaccaggt agtggccgcc gtccttcagc ttcagggcct ggtggatctc 481 gcccttcagc acgccgtcgc gggggtacag gcgctcggtg gaggcctccc agcccatggt 541 cttcttctgc attacggggc cgtcgggggg gaagttggtg ccgcgcatct tcaccttgta 601 gatcagcgtg ccgtcctgca gggaggagtc ctgggtcacg gtcaccagac cgccgtcctc 661 gaagttcatc acgcgctccc acttgaagcc ctcggggaag gacagcttct tgtaatcggg 721 gatgtcggcg gggtgcttca cgtacgcctt ggagccgtac atgaactggg gggacaggat 781 gtcccaggcg aagggcaggg ggccgccctt ggtcaccttc agcttggcgg tctgggtgcc 841 ctcgtagggg cggccctcgc cctcgccctc gatctcgaac tcgtggccgt tcatggagcc 901 ctccatgcgc accttgaagc gcatgaactc tttgatgacg gccatgttgt tgtcctcgga 961 ggaggcggtg ccggagctgc cgctgccggt gctgccggtg ccatgcccca ggaacaggtg 1021 gtggcggccc tcggagcgct cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt 1081 gatgtccagc ttggtgtcca cgtagtagta gccgggcagt tgcacgggct tcttggccat 1141 gtagatggtc ttgaactcca ccaggtagtg gccgccgtcc ttcagcttca gggcctggtg 1201 gatctcgccc ttcagcacgc cgtcgcgggg gtacaggcgc tcggtggagg cctcccagcc 1261 catggtcttc ttctgcatta cggggccgtc gggggggaag ttggtgccgc gcatcttcac 1321 cttgtagatc agcgtgccgt cctgcaggga ggagtcctgg gtcacggtca ccagaccgcc 1381 gtcctcgaag ttcatcacgc gctcccactt gaagccctcg gggaaggaca gcttcttgta 1441 atcggggatg tcggcggggt gcttcacgta cgccttggag ccgtacatga actgggggga 1501 caggatgtcc caggcgaagg gcagggggcc gcccttggtc accttcagct tggcggtctg 1561 ggtgccctcg taggggcggc cctcgccctc gccctcgatc tcgaactcgt ggccgttcat 1621 ggagccctcc atgcgcacct tgaagcgcat gaactctttg atgacctcct cgcccttgct 1681 caccatatcg atgctgctct cgtacttgcc ggtgcccttg ttcacggcgc cggcctcggc 1741 ctcgtcctcc accagggtct ccacctcgat ctcggtgccg ccgatgttca gcttggtggt 1801 ctttctgatg tcgccgtgga tcaggatgtg ctcgtggatc agcactctca ggtagtggcc 1861 cagcaggccc cagcagttct tgctcatcag gtcgatgatg gtgtggccca cggtgctgcc 1921 gtacacgctc agcacgccga agccctcggg gcccaggatg aacaggatgg ggaacatgcc 1981 ccagctcacg aagaacagcc aggccatgcc ggtcaccacc tgtctgcatc tgcccttggg 2041 cacggtgtgg taggcctcga tgtacacctt ggcggcgttg aagaaggtgt agatgccgta 2101 gcacaggccc atcaggaaga agatcactct cacgtagccc ttgctcaggg cggcggtggt 2161 gccccacacg atggtgccga tgtcgctcac cagcaggccc atggttctct tgttgtagtc 2221 gttggccagg ccggtcaggt tgctcaggtg gatcaggacc acggggcagg tcagcagcca 2281 ctcggcgtat ctcagccaca cggtcttgtt gccgttgctg ctgtagatca cggcgggctc 2341 gtcgaactcg tggaagtact ctatgatgaa cttgatcatc tcgatggtgg ccacgtagat 2401 ctcctcccag ccgcaggtac tcttccaggt ctggtagccg tagaacatca ggcacagggc 2461 gctcagggcg aaggtgatcc actgcaggat gttggcggcc agcttctcgg cgttggtgcc 2521 gttactcttc agccaggcca ggcagaagca ctggccgttg ttggggatgc agatcacgct 2581 gccgttgttc tccagggtgt agctggtctg gaacagcatt ctctcgtggg ctctgtggaa 2641 cacgtagtcg gggccgtcct gggtggccac gggcacggtg gcgtcgctgc cggtgctggc 2701 gccggcgctg ccggcggcca gggccacggc cagggccagg gccagcagcc agggtcttct 2761 gctcaccatg gtcaccggtg gtaccgaatt ctagtataac ttcgtatagg atactttata 2821 cgaagttatc attgggattc ttcctatttt gatccaagca tcaccatcga ccctctagtc 2881 cagatctcac catcgaccca taacttcgta tagcatacat tatacgaagt tatgtccctc 2941 gaagaggttc gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg 3001 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc 3061 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta 3121 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt 3181 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca 3241 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc 3301 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt 3361 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg 3421 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg 3481 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat 3541 ctccctttgg gccgcctccc cgcatcgata ccgtcgaccc gggcggccgc ttcgagcaga 3601 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg 3661 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa 3721 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga 3781 ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg atcttcctag 3841 agcatggcta cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag 3901 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa 3961 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct 4021 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 4081 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 4141 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 4201 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 4261 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 4321 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 4381 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 4441 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 4501 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 4561 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 4621 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 4681 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 4741 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 4801 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 4861 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 4921 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 4981 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 5041 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 5101 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 5161 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 5221 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 5281 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 5341 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 5401 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 5461 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 5521 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 5581 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 5641 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 5701 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 5761 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 5821 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 5881 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 5941 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 6001 acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc 6061 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc 6121 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac 6181 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca 6241 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg 6301 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag 6361 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc 6421 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg 6481 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctgcgcgct 6541 cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg 6601 gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc 6661 ttgtagttaa tgattaaccc gccatgctac ttatctacgt agccatgctc taggaagatc 6721 gtaccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat 6781 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat 6841 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat 6901 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc 6961 gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc 7021 tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg 7081 gggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg gggcggggcg 7141 aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg 7201 gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct 7261 gcgacgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct 7321 ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg 7381 taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg aaagccttga 7441 ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggctgtccg cggggggacg 7501 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct //