LOCUS Exported 6516 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Cre expression. ACCESSION . VERSION . KEYWORDS pAAV-Syn-iCre SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6516) TITLE AAV expression plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 6516) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6516 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 48..350 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" regulatory 1383..1392 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1395..1415 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 2451..2477 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" misc_feature 2503..3091 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2556..2576) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2974..2985) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3116..3237 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3127..3146) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3181..3200 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 3337..3466 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" rep_origin complement(3528..4116) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3608..3627) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4287..5147) /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 4910..4929 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5148..5252) /gene="bla" /label=AmpR promoter primer_bind 5320..5338 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(5376..5398) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 5498..5517 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" rep_origin complement(5534..5989) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5671..5692) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5883..5902 /label=F1ori-R /note="F1 origin, reverse primer" repeat_region 6064..6204 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 6075..6204 /label=AAV2 ITR ORIGIN 1 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt 61 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg 121 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat 181 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct 241 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg 301 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat tcagaaccac 361 ggacagcacc gaattcaatg tgtaatgtgt gctcaatatg ctcatgtgtg ttaccctgag 421 cttctgtgtc tacatatata cctggatgcc tgtgtgttct gtgatgtaca tatatattct 481 gtctttcctt ccttttctat ttgtgttatt ccatgtgttc tttcagattc tcaccaccaa 541 gggcaaggat atgttaacta cccaagtgtc cacctccgcc tgtctggtga tgtttacgcc 601 acccccgtgc tcttttcttt gcccgacaga gttgttatag gagatgtctc cccgggaaca 661 ctgcaggaag gagaatttct acatttatgt tcccctctga gtgtgcttct atccccaaaa 721 tgccttcaaa ggtgaaaatc aacactggaa acccaagtat ctgggaaggg caagagtgtg 781 taagtgcaag ttagcctaag gaataggaag aggttggtaa acagggtagg atcgtgggag 841 ggagtttcgt tactacaggt ccggaccctc aggacaagaa ccccaccccc actccccaaa 901 ttgcgcatcc cccgccccca tcagaggggg aggggaagag gttgcggcgc ggcgcatgcg 961 cactgtcgga ttcagcaccg cggtcagagc cttcgcctcc gctgccggcg cgcaccacca 1021 cctccccagc accaaaggct gactgacgtc actcactagc cctccccaaa ctccccttcc 1081 tcgccgcctt ggtcgcgtcc atgctgccgt gagtccagtc ggaccgcacc acgagaggtg 1141 caagataggg aggtgcgggc gcgaccatac gctctgcggc ggcagagcct cggcgctgcc 1201 tcagtctgca gcgggcagca gaggagtcgc gtcgtgccag agagcaccgc cgtgctcctg 1261 agcccctcgc gctccgcccc cgcggcccac agacccactg ccccttggat cccctgcccc 1321 acgccagagc ctctgctaac catgttcatg ccttcttctt tttcctacag gagctcgcca 1381 ccgccaccat ggtgcccaag aagaagagga aagtctccaa cctgctgact gtgcaccaaa 1441 acctgcctgc cctccctgtg gatgccacct ctgatgaagt caggaagaac ctgatggaca 1501 tgttcaggga caggcaggcc ttctctgaac acacctggaa gatgctcctg tctgtgtgca 1561 gatcctgggc tgcctggtgc aagctgaaca acaggaaatg gttccctgct gaacctgagg 1621 atgtgaggga ctacctcctg tacctgcaag ccagaggcct ggctgtgaag accatccaac 1681 agcacctggg ccagctcaac atgctgcaca ggagatctgg cctgcctcgc ccttctgact 1741 ccaatgctgt gtccctggtg atgaggagaa tcagaaagga gaatgtggat gctggggaga 1801 gagccaagca ggccctggcc tttgaacgca ctgactttga ccaagtcaga tccctgatgg 1861 agaactctga cagatgccag gacatcagga acctggcctt cctgggcatt gcctacaaca 1921 ccctgctgcg cattgccgaa attgccagaa tcagagtgaa ggacatctcc cgcaccgatg 1981 gtgggagaat gctgatccac attggcagga ccaagaccct ggtgtccaca gctggtgtgg 2041 agaaggccct gtccctgggg gttaccaagc tggtggagag atggatctct gtgtctggtg 2101 tggctgatga ccccaacaac tacctgttct gccgggtcag aaagaatggt gtggctgccc 2161 cttctgccac ctcccaactg tccacccggg ccctggaagg gatctttgag gccacccacc 2221 gcctgatcta tggtgccaag gatgactctg ggcagagata cctggcctgg tctggccact 2281 ctgccagagt gggtgctgcc agggacatgg ccagggctgg tgtgtccatc cctgaaatca 2341 tgcaggctgg tggctggacc aatgtgaaca ttgtgatgaa ctacatcaga aacctggact 2401 ctgagactgg ggccatggtg aggctgctcg aggatgggga cagcggcgga tacccttacg 2461 acgtgcctga ctacgcctga ctgcagaacg gccgttatcg ataatcaacc tctggattac 2521 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 2581 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 2641 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 2701 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 2761 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 2821 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 2881 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg 2941 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 3001 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg 3061 agtcggatct ccctttgggc cgcctccccg catcgattgc ggccgtcaga catgataaga 3121 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt 3181 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac 3241 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa 3301 agcaagtaaa acctctacaa atgtggtagc ggccgcagga acccctagtg atggagttgg 3361 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac 3421 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggacat gtgagcaaaa 3481 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 3541 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 3601 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 3661 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 3721 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 3781 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 3841 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 3901 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 3961 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 4021 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 4081 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 4141 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 4201 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 4261 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 4321 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg 4381 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca 4441 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 4501 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 4561 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca 4621 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 4681 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 4741 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 4801 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 4861 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg 4921 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 4981 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 5041 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 5101 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 5161 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 5221 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac 5281 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc 5341 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag 5401 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca 5461 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg 5521 agagtgcacc ataaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt 5581 taaatcagct cattttttaa ccaatagacc gaaatcggca aaatccctta taaatcaaaa 5641 gaatagcccg agatagagtt gagtgttgtt ccagtttgga acaagagtcc actattaaag 5701 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt 5761 gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac 5821 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag 5881 gaagggaaga aagcgaaagg agcgggcgct aaggcgctgg caagtgtagc ggtcacgctg 5941 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgta ctatggttgc 6001 tttgacgtat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc 6061 gcccctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggca aagcccgggc 6121 gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg 6181 ccaactccat cactaggggt tcctgcggcc gcggtacctt aattaaggga tgtaattacg 6241 tccctaaccc gctagggggc agcagcgagc cgaacggggc tccgctccgg tccggcgcta 6301 accccgcatc cccgaggggg gatgtaatta cgtccctaac ccgctagggg gcagcagcga 6361 gccgaacggg gctccgctcc ggtccggcgc taaccccgca tccccgaggg gggatgtaat 6421 tacgtcccta acccgctagg gggcagcagc gagccgaacg gggctccgct ccggtccggc 6481 gctaaccccg catccccgag ggtcgactat taatag //