LOCUS Exported 7336 bp ds-DNA circular SYN 25-MAY-2021 DEFINITION AAV packaging plasmid expressing Rep/Cap genes. ACCESSION . VERSION . KEYWORDS pAAV2/8 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7336) TITLE AAV packaging plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 7336) AUTHORS . TITLE Direct Submission JOURNAL Exported May 25, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7336 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 3941..3952 /codon_start=1 /product="WELQut protease recognition and cleavage site" /label=WELQut site /translation="WELQ" primer_bind complement(4404..4424) /label=T3 /note="T3 promoter, forward primer" promoter complement(4404..4422) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(4443..4459) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4443..4459) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4456..4478) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4467..4483 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(4491..4521) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4536..4557 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4674..4691) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4845..5433) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4925..4944) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5604..6464) /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 6227..6246 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6465..6569) /gene="bla" /label=AmpR promoter rep_origin complement(6595..7050) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6732..6753) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 6944..6963 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7177..7199 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 7191..7208 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 7192..7208 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 7218..7237 /label=T7 /note="T7 promoter, forward primer" promoter 7218..7236 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind 7265..7281 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 7266..7282 /label=KS primer /note="common sequencing primer, one of multiple similar variants" ORIGIN 1 atgccggggt tttacgagat tgtgattaag gtccccagcg accttgacga gcatctgccc 61 ggcatttctg acagctttgt gaactgggtg gccgagaagg aatgggagtt gccgccagat 121 tctgacatgg atctgaatct gattgagcag gcacccctga ccgtggccga gaagctgcag 181 cgcgactttc tgacggaatg gcgccgtgtg agtaaggccc cggaggctct tttctttgtg 241 caatttgaga agggagagag ctacttccac atgcacgtgc tcgtggaaac caccggggtg 301 aaatccatgg ttttgggacg tttcctgagt cagattcgcg aaaaactgat tcagagaatt 361 taccgcggga tcgagccgac tttgccaaac tggttcgcgg tcacaaagac cagaaatggc 421 gccggaggcg ggaacaaggt ggtggatgag tgctacatcc ccaattactt gctccccaaa 481 acccagcctg agctccagtg ggcgtggact aatatggaac agtatttaag cgcctgtttg 541 aatctcacgg agcgtaaacg gttggtggcg cagcatctga cgcacgtgtc gcagacgcag 601 gagcagaaca aagagaatca gaatcccaat tctgatgcgc cggtgatcag atcaaaaact 661 tcagccaggt acatggagct ggtcgggtgg ctcgtggaca aggggattac ctcggagaag 721 cagtggatcc aggaggacca ggcctcatac atctccttca atgcggcctc caactcgcgg 781 tcccaaatca aggctgcctt ggacaatgcg ggaaagatta tgagcctgac taaaaccgcc 841 cccgactacc tggtgggcca gcagcccgtg gaggacattt ccagcaatcg gatttataaa 901 attttggaac taaacgggta cgatccccaa tatgcggctt ccgtctttct gggatgggcc 961 acgaaaaagt tcggcaagag gaacaccatc tggctgtttg ggcctgcaac taccgggaag 1021 accaacatcg cggaggccat agcccacact gtgcccttct acgggtgcgt aaactggacc 1081 aatgagaact ttcccttcaa cgactgtgtc gacaagatgg tgatctggtg ggaggagggg 1141 aagatgaccg ccaaggtcgt ggagtcggcc aaagccattc tcggaggaag caaggtgcgc 1201 gtggaccaga aatgcaagtc ctcggcccag atagacccga ctcccgtgat cgtcacctcc 1261 aacaccaaca tgtgcgccgt gattgacggg aactcaacga ccttcgaaca ccagcagccg 1321 ttgcaagacc ggatgttcaa atttgaactc acccgccgtc tggatcatga ctttgggaag 1381 gtcaccaagc aggaagtcaa agactttttc cggtgggcaa aggatcacgt ggttgaggtg 1441 gagcatgaat tctacgtcaa aaagggtgga gccaagaaaa gacccgcccc cagtgacgca 1501 gatataagtg agcccaaacg ggtgcgcgag tcagttgcgc agccatcgac gtcagacgcg 1561 gaagcttcga tcaactacgc agacaggtac caaaacaaat gttctcgtca cgtgggcatg 1621 aatctgatgc tgtttccctg cagacaatgc gagagaatga atcagaattc aaatatctgc 1681 ttcactcacg gacagaaaga ctgtttagag tgctttcccg tgtcagaatc tcaacccgtt 1741 tctgtcgtca aaaaggcgta tcagaaactg tgctacattc atcatatcat gggaaaggtg 1801 ccagacgctt gcactgcctg cgatctggtc aatgtggatt tggatgactg catctttgaa 1861 caataaatga tttaaatcag gtatggctgc cgatggttat cttccagatt ggctcgagga 1921 caacctctct gagggcattc gcgagtggtg ggcgctgaaa cctggagccc cgaagcccaa 1981 agccaaccag caaaagcagg acgacggccg gggtctggtg cttcctggct acaagtacct 2041 cggacccttc aacggactcg acaaggggga gcccgtcaac gcggcggacg cagcggccct 2101 cgagcacgac aaggcctacg accagcagct gcaggcgggt gacaatccgt acctgcggta 2161 taaccacgcc gacgccgagt ttcaggagcg tctgcaagaa gatacgtctt ttgggggcaa 2221 cctcgggcga gcagtcttcc aggccaagaa gcgggttctc gaacctctcg gtctggttga 2281 ggaaggcgct aagacggctc ctggaaagaa gagaccggta gagccatcac cccagcgttc 2341 tccagactcc tctacgggca tcggcaagaa aggccaacag cccgccagaa aaagactcaa 2401 ttttggtcag actggcgact cagagtcagt tccagaccct caacctctcg gagaacctcc 2461 agcagcgccc tctggtgtgg gacctaatac aatggctgca ggcggtggcg caccaatggc 2521 agacaataac gaaggcgccg acggagtggg tagttcctcg ggaaattggc attgcgattc 2581 cacatggctg ggcgacagag tcatcaccac cagcacccga acctgggccc tgcccaccta 2641 caacaaccac ctctacaagc aaatctccaa cgggacatcg ggaggagcca ccaacgacaa 2701 cacctacttc ggctacagca ccccctgggg gtattttgac tttaacagat tccactgcca 2761 cttttcacca cgtgactggc agcgactcat caacaacaac tggggattcc ggcccaagag 2821 actcagcttc aagctcttca acatccaggt caaggaggtc acgcagaatg aaggcaccaa 2881 gaccatcgcc aataacctca ccagcaccat ccaggtgttt acggactcgg agtaccagct 2941 gccgtacgtt ctcggctctg cccaccaggg ctgcctgcct ccgttcccgg cggacgtgtt 3001 catgattccc cagtacggct acctaacact caacaacggt agtcaggccg tgggacgctc 3061 ctccttctac tgcctggaat actttccttc gcagatgctg agaaccggca acaacttcca 3121 gtttacttac accttcgagg acgtgccttt ccacagcagc tacgcccaca gccagagctt 3181 ggaccggctg atgaatcctc tgattgacca gtacctgtac tacttgtctc ggactcaaac 3241 aacaggaggc acggcaaata cgcagactct gggcttcagc caaggtgggc ctaatacaat 3301 ggccaatcag gcaaagaact ggctgccagg accctgttac cgccaacaac gcgtctcaac 3361 gacaaccggg caaaacaaca atagcaactt tgcctggact gctgggacca aataccatct 3421 gaatggaaga aattcattgg ctaatcctgg catcgctatg gcaacacaca aagacgacga 3481 ggagcgtttt tttcccagta acgggatcct gatttttggc aaacaaaatg ctgccagaga 3541 caatgcggat tacagcgatg tcatgctcac cagcgaggaa gaaatcaaaa ccactaaccc 3601 tgtggctaca gaggaatacg gtatcgtggc agataacttg cagcagcaaa acacggctcc 3661 tcaaattgga actgtcaaca gccagggggc cttacccggt atggtctggc agaaccggga 3721 cgtgtacctg cagggtccca tctgggccaa gattcctcac acggacggca acttccaccc 3781 gtctccgctg atgggcggct ttggcctgaa acatcctccg cctcagatcc tgatcaagaa 3841 cacgcctgta cctgcggatc ctccgaccac cttcaaccag tcaaagctga actctttcat 3901 cacgcaatac agcaccggac aggtcagcgt ggaaattgaa tgggagctgc agaaggaaaa 3961 cagcaagcgc tggaaccccg agatccagta cacctccaac tactacaaat ctacaagtgt 4021 ggactttgct gttaatacag aaggcgtgta ctctgaaccc cgccccattg gcacccgtta 4081 cctcacccgt aatctgtaat tgcctgttaa tcaataaacc ggttgattcg tttcagttga 4141 actttggtct ctgcgaaggg cgaattcgtt taaacctgca ggactagagg tcctgtatta 4201 gaggtcacgt gagtgttttg cgacattttg cgacaccatg tggtcacgct gggtatttaa 4261 gcccgagtga gcacgcaggg tctccatttt gaagcgggag gtttgaacgc gcagccgcca 4321 agccgaattc tgcagatatc catcacactg gcggccgctc gactagagcg gccgccaccg 4381 cggtggagct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca 4441 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga 4501 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt 4561 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga 4621 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc 4681 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg 4741 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc 4801 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 4861 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 4921 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 4981 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat 5041 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 5101 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 5161 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 5221 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 5281 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 5341 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag 5401 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg 5461 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa 5521 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata 5581 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg 5641 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata 5701 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg 5761 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct 5821 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt 5881 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc 5941 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga 6001 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt 6061 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc 6121 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa 6181 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca 6241 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca 6301 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct 6361 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc 6421 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa 6481 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt 6541 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg 6601 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta 6661 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt 6721 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca 6781 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa 6841 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat 6901 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag 6961 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg 7021 ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt 7081 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg 7141 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga 7201 cggccagtga gcgcgcgtaa tacgactcac tatagggcga attgggtacc gggccccccc 7261 tcgatcgagg tcgacggtat cgggggagct cgcagggtct ccattttgaa gcgggaggtt 7321 tgaacgcgca gccgcc //