LOCUS Exported 6522 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DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6522) TITLE AAV expression plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 6522) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6522 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(11..28) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(182..770) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(262..281) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(941..1801) /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 1564..1583 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1802..1906) /gene="bla" /label=AmpR promoter rep_origin complement(1932..2387) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2069..2090) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2281..2300 /label=F1ori-R /note="F1 origin, reverse primer" repeat_region 2459..2588 /label=AAV2 ITR promoter 2901..3178 /label=chicken beta-actin promoter primer_bind 3458..3480 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 3542..3561 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" protein_bind complement(3596..3629) /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(3692..3725) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." regulatory 5197..5206 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind 5245..5278 /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 5349..5382 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 5424..6012 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5477..5497) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5895..5906) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 6055..6176 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6066..6085) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 6120..6139 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 6338..6467 /label=AAV2 ITR ORIGIN 1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta 1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga 2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa 2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga 2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc 2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga 2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg 2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt 2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct 2461 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg 2521 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag 2581 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag 2641 gaagatcgta ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac 2701 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca 2761 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg 2821 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt 2881 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc 2941 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat 3001 gggggcgggg gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg 3061 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc 3121 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg 3181 agtcgctgcg acgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc 3241 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc 3301 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa 3361 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ctgtccgcgg 3421 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc 3481 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg 3541 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattgga tccccataac 3601 ttcgtataaa gtatcctata cgaagttata tcaaaatagg aagaccaatg cttcaccatc 3661 gacccgaatt gccaagcatc accatcgacc cataacttcg tataatgtat gctatacgaa 3721 gttatactag tgctagagct agctgatcac ttaaggttta aacaccggtt tatttataca 3781 gttcgtccat accatacaga aacagatgat ggcgaccttc agaacgctcg tactgttcaa 3841 caatagtata atcctcgttg tgggaggtga tgtccagttt agtgtcaacg tagtaatagc 3901 ccggcagttg tacaggcttt tttgccatgt agatagtttt gaactcaacc agatagtgac 3961 caccatcctt cagcttcagg gcctggtgaa tctcgccttt cagcacaccg tcgcgcgggt 4021 acagacgttc ggtgctagct tcccagccca tagttttctt ctgcattact ggaccgtcag 4081 gcgggaagtt agtgccacgc attttcactt tgtaaatcag agtaccgtct tgcaggctgc 4141 tgtcctgcgt aacagtgacc aggccgccat cttcgaaatt cataacacgt tcccatttga 4201 agccctctgg gaaggacagc tttttgtaat ccgggatgtc tgccgggtgt tttacgtagg 4261 ctttggaacc gtacatgaac tgcggagaca ggatatccca cgcaaaaggc agtgggccac 4321 ctttggtaac tttcagcttt gcggtctgag tgccctcgta tgggcgaccc tcgccctcac 4381 cttcaatctc gaactcatgg ccgttcatag aaccttccat acgtactttg aacctcatga 4441 actctttgat caccgccatg ttgttgtctt cggaagacgc agtgccagag gaaccagaac 4501 ccgtagaacc agtaccgtgg cccaggaaca ggtggtgacg gccctcgcta cgttcatact 4561 gttcgacgat ggtataatct tcgttgtgag acgtgatatc cagtttggtg tccacgtagt 4621 agtaacctgg cagttgaacc ggctttttag ccatatagat cgtcttaaat tcaaccaggt 4681 agtgaccgcc gtccttcagt ttcagcgcct ggtggatctc acctttcaga acaccgtcac 4741 gcggatacag gcgctcggta gatgcttccc atcccatcgt ctttttctgc atcaccgggc 4801 catccggcgg aaagttcgta ccacgcatct tcaccttgta gatcagggtg ccatcttgca 4861 gagagctatc ctgggtgacg gtaaccaggc caccgtcctc aaagttcatt acacgctccc 4921 acttgaaacc ctccgggaaa gacagcttct tgtagtccgg gatatcggct gggtgtttga 4981 cgtaggcctt ggaaccatac ataaactgcg gggacaggat gtcccacgcg aacggcagag 5041 gaccgccttt ggttactttc agtttagcgg tctgcgtacc ctcgtacgga cgaccttcgc 5101 cttcgccctc gatctcaaac tcatgaccgt tcatgctacc ctccatgcgt actttaaaac 5161 gcataaactc tttaatcacc tcttcaccct tgctaaccat ggtggcctcg agtcgcgaac 5221 gcgtggcgcg ccggtaccac tagtataact tcgtatagga tactttatac gaagttatca 5281 ttgggattct tcctattttg atccaagcat caccatcgac cctctagtcc agatctcacc 5341 atcgacccat aacttcgtat agcatacatt atacgaagtt atgtccctcg aagaggttcg 5401 aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga 5461 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt 5521 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt 5581 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg 5641 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg 5701 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc 5761 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat 5821 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct 5881 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg 5941 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg 6001 ccgcctcccc gcatcgatac cgtcgacccg ggcggccgct tcgagcagac atgataagat 6061 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg 6121 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca 6181 acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa 6241 gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tcttcctaga gcatggctac 6301 gtagataagt agcatggcgg gttaatcatt aactacaagg aacccctagt gatggagttg 6361 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga 6421 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cattaatgaa 6481 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ct //