LOCUS Exported 6519 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Cre dependent TdTomato expression.. ACCESSION . VERSION . KEYWORDS pAAV-CAG-Flex-Rev-TdTomato SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6519) TITLE AAV expression plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 6519) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6519 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..278 /label=chicken beta-actin promoter primer_bind 557..579 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 641..660 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" protein_bind complement(695..728) /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(791..824) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." regulatory 2296..2305 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind 2344..2377 /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 2448..2481 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 2523..3111 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2576..2596) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2994..3005) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3154..3275 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3165..3184) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3219..3238 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 3437..3566 /label=AAV2 ITR primer_bind complement(3632..3649) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3803..4391) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3883..3902) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4562..5422) /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 5185..5204 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5423..5527) /gene="bla" /label=AmpR promoter rep_origin complement(5553..6008) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5690..5711) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5902..5921 /label=F1ori-R /note="F1 origin, reverse primer" repeat_region 6080..6209 /label=AAV2 ITR ORIGIN 1 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa 61 ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg 121 ggcgcgcgcc rggsggggsg gggsggggsg rggggsgggg sggggsgagg cggagaggtg 181 cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc 241 ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc 301 gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt 361 tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg 421 tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg 481 gccctttgtg cggggggagc ggctcggggc tgtccgcggg gggacggctg ccttcggggg 541 ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag agcctctgct 601 aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg 661 ctgtctcatc attttggcaa agaattggat ccccataact tcgtataaag tatcctatac 721 gaagttatat caaaatagga agaccaatgc ttcaccatcg acccgaattg ccaagcatca 781 ccatcgaccc ataacttcgt ataatgtatg ctatacgaag ttatactagt gctagagcta 841 gctgatcact taaggtttaa acaccggttt atttatacag ttcgtccata ccatacagaa 901 acagatgatg gcgaccttca gaacgctcgt actgttcaac aatagtataa tcctcgttgt 961 gggaggtgat gtccagttta gtgtcaacgt agtaatagcc cggcagttgt acaggctttt 1021 ttgccatgta gatagttttg aactcaacca gatagtgacc accatccttc agcttcaggg 1081 cctggtgaat ctcgcctttc agcacaccgt cgcgcgggta cagacgttcg gtgctagctt 1141 cccagcccat agttttcttc tgcattactg gaccgtcagg cgggaagtta gtgccacgca 1201 ttttcacttt gtaaatcaga gtaccgtctt gcaggctgct gtcctgcgta acagtgacca 1261 ggccgccatc ttcgaaattc ataacacgtt cccatttgaa gccctctggg aaggacagct 1321 ttttgtaatc cgggatgtct gccgggtgtt ttacgtaggc tttggaaccg tacatgaact 1381 gcggagacag gatatcccac gcaaaaggca gtgggccacc tttggtaact ttcagctttg 1441 cggtctgagt gccctcgtat gggcgaccct cgccctcacc ttcaatctcg aactcatggc 1501 cgttcataga accttccata cgtactttga acctcatgaa ctctttgatc accgccatgt 1561 tgttgtcttc ggaagacgca gtgccagagg aaccagaacc cgtagaacca gtaccgtggc 1621 ccaggaacag gtggtgacgg ccctcgctac gttcatactg ttcgacgatg gtataatctt 1681 cgttgtgaga cgtgatatcc agtttggtgt ccacgtagta gtaacctggc agttgaaccg 1741 gctttttagc catatagatc gtcttaaatt caaccaggta gtgaccgccg tccttcagtt 1801 tcagcgcctg gtggatctca cctttcagaa caccgtcacg cggatacagg cgctcggtag 1861 atgcttccca tcccatcgtc tttttctgca tcaccgggcc atccggcgga aagttcgtac 1921 cacgcatctt caccttgtag atcagggtgc catcttgcag agagctatcc tgggtgacgg 1981 taaccaggcc accgtcctca aagttcatta cacgctccca cttgaaaccc tccgggaaag 2041 acagcttctt gtagtccggg atatcggctg ggtgtttgac gtaggccttg gaaccataca 2101 taaactgcgg ggacaggatg tcccacgcga acggcagagg accgcctttg gttactttca 2161 gtttagcggt ctgcgtaccc tcgtacggac gaccttcgcc ttcgccctcg atctcaaact 2221 catgaccgtt catgctaccc tccatgcgta ctttaaaacg cataaactct ttaatcacct 2281 cttcaccctt gctaaccatg gtggcctcga gtcgcgaacg cgtggcgcgc cggtaccact 2341 agtataactt cgtataggat actttatacg aagttatcat tgggattctt cctattttga 2401 tccaagcatc accatcgacc ctctagtcca gatctcacca tcgacccata acttcgtata 2461 gcatacatta tacgaagtta tgtccctcga agaggttcga attcgatatc aagcttatcg 2521 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg 2581 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc 2641 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt 2701 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca 2761 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc 2821 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc 2881 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc 2941 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc 3001 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc 3061 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc 3121 gtcgacccgg gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa 3181 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt 3241 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta 3301 tgtttcaggt tcagggggag atgtgggagg ttttttaaag caagtaaaac ctctacaaat 3361 gtggtaaaat cgataaggat cttcctagag catggctacg tagataagta gcatggcggg 3421 ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct 3481 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg 3541 gcctcagtga gcgagcgagc gcgcagctgc attaatgaat cggccaacgc gcggggagag 3601 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 3661 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 3721 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 3781 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 3841 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 3901 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 3961 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 4021 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 4081 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 4141 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 4201 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 4261 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 4321 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa 4381 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa 4441 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt 4501 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca 4561 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 4621 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc 4681 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa 4741 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc 4801 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca 4861 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat 4921 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag 4981 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac 5041 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt 5101 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt 5161 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc 5221 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat 5281 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca 5341 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga 5401 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg 5461 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg 5521 ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa 5581 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa 5641 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa 5701 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca 5761 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg 5821 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc 5881 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc 5941 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca 6001 gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc 6061 ctcttcgcta ttacgccagc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc 6121 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg 6181 agtggccaac tccatcacta ggggttcctt gtagttaatg attaacccgc catgctactt 6241 atctacgtag ccatgctcta ggaagagtac cattgacgtc aataatgacg tatgttccca 6301 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg 6361 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg 6421 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt 6481 ggcagtacat ctacgtatta gtcatcgcta ttaccatgg //