LOCUS Exported 8469 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION non-standard AAV2 rep-AAV-PHP.V2 cap plasmid with AAV cap expression controlled by a tTA-TRE amplification system. ACCESSION . VERSION . KEYWORDS pUCmini-iCAP-PHP.V2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8469) AUTHORS Kumar SR, Miles TF, Chen X, Brown D, Dobreva T, Huang Q, Ding X, Luo Y, Einarsson PH, Greenbaum A, Jang MJ, Deverman BE, Gradinaru V. TITLE Multiplexed Cre-dependent selection yields systemic AAVs for targeting distinct brain cell types. JOURNAL Nat Methods (2020). REFERENCE 2 (bases 1 to 8469) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8469 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 99..121 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 113..130 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 114..130 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 140..159 /label=T7 /note="T7 promoter, forward primer" promoter 140..158 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 2748..2858 /codon_start=1 /product="3 tandem repeats of the minimal activation domain of herpes simplex virus VP16 (Cheng et al., 2013)" /label=VP48 /translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML" promoter complement(2897..2915) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2898..2915) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(2935..2952) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2941..3165 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" protein_bind 3227..3245 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 3264..3282 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 3301..3319 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 3338..3356 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 3375..3393 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 3412..3430 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" CDS 6147..6158 /codon_start=1 /product="WELQut protease recognition and cleavage site" /label=WELQut site /translation="WELQ" rep_origin complement(6714..7302) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6794..6813) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7473..8333) /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 8096..8115 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8334..8438) /gene="bla" /label=AmpR promoter ORIGIN 1 ccaatgatac gcgtcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg 61 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac 121 gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta catcgacggt 181 atcgggggag ctcgcagggt ctccattttg aagcgggagg tttgaacgcg cagccgccat 241 gccggggttt tacgagattg tgattaaggt ccccagcgac cttgacgagc atctgcccgg 301 catttctgac agctttgtga actgggtggc cgagaaggaa tgggagttgc cgccagattc 361 tgacatggat ctgaatctga ttgagcaggc acccctgacc gtggccgaga agctgcagcg 421 cgactttctg acggaatggc gccgtgtgag taaggccccg gaggctcttt tctttgtgca 481 atttgagaag ggagagagct acttccacat gcacgtgctc gtggaaacca ccggggtgaa 541 atccatggtt ttgggacgtt tcctgagtca gattcgcgaa aaactgattc agagaattta 601 ccgcgggatc gagccgactt tgccaaactg gttcgcggtc acaaagacca gaaatggcgc 661 cggaggcggg aacaaggtgg tggatgagtg ctacatcccc aattacttgc tccccaaaac 721 ccagcctgag ctccagtggg cgtggactaa tatggaacag tatttaagcg cctgtttgaa 781 tctcacggag cgtaaacggt tggtggcgca gcatctgacg cacgtgtcgc agacgcagga 841 gcagaacaaa gagaatcaga atcccaattc tgatgcgccg gtgatcagat caaaaacttc 901 agccaggtac atggagctgg tcgggtggct cgtggacaag gggattacct cggagaagca 961 gtggatccag gaggaccagg cctcatacat ctccttcaat gcggcctcca actcgcggtc 1021 ccaaatcaag gctgccttgg acaatgcggg aaagattatg agcctgacta aaaccgcccc 1081 cgactacctg gtgggccagc agcccgtgga ggacatttcc agcaatcgga tttataaaat 1141 tttggaacta aacgggtacg atccccaata tgcggcttcc gtctttctgg gatgggccac 1201 gaaaaagttc ggcaagagga acaccatctg gctgtttggg cctgcaacta ccgggaagac 1261 caacatcgcg gaggccatag cccacactgt gcccttctac gggtgcgtaa actggaccaa 1321 tgagaacttt cccttcaacg actgtgtgga caagatggtg atctggtggg aggaggggaa 1381 gatgaccgcc aaggtcgtgg agtcggccaa agccattctc ggaggaagca aggtgcgcgt 1441 ggaccagaaa tgcaagtcct cggcccagat agacccgact cccgtgatcg tcacctccaa 1501 caccaatatg tgcgccgtga ttgacgggaa ctcaacgacc ttcgaacacc agcagccgtt 1561 gcaagaccgg atgttcaaat ttgaactcac ccgccgtctg gatcatgact ttgggaaggt 1621 caccaagcag gaagtcaaag actttttccg gtgggcaaag gatcacgtgg ttgaggtgga 1681 gcatgaattc tacgtcaaaa agggtggagc caagaaaaga cccgccccca gtgacgcaga 1741 tataagtgag cccaaacggg tgcgcgagtc agttgcgcag ccatcgacgt cagacgcgga 1801 agcttcgatc aactacgcgg acaggtacca aaacaaatgt tctcgtcacg tgggcatgaa 1861 tctgatgctg tttccctgca gacaatgcga gagactgaat cagaattcaa atatctgctt 1921 cactcacggt gtcaaagact gtttagagtg ctttcccgtg tcagaatctc aacccgtttc 1981 tgtcgtcaaa aaggcgtatc agaaactgtg ctacattcat cacatcatgg gaaaggtgcc 2041 agacgcttgc actgcttgcg acctggtcaa tgtggacttg gatgactgtg tttctgaaca 2101 ataaatgact taaaccaggt atgagtcggc tggataaatc taaagtcata aacggcgctc 2161 tggaattact caatgaagtc ggtatcgaag gcctgacgac aaggaaactc gctcaaaagc 2221 tgggagttga gcagcctacc ctgtactggc acgtgaagaa caagcgggcc ctgctcgatg 2281 ccctggccat cgagatgctg gacaggcatc atacccactt ctgccccctg gaaggcgagt 2341 catggcaaga ctttctgcgg aacaacgcca agtcattccg ctgtgctctc ctctcacatc 2401 gcgacggggc taaagtgcat ctcggcaccc gcccaacaga gaaacagtac gaaaccctgg 2461 aaaatcagct cgcgttcctg tgtcagcaag gcttctccct ggagaacgca ctgtacgctc 2521 tgtccgccgt gggccacttt acactgggct gcgtattgga ggaacaggag catcaagtag 2581 caaaagagga aagagagaca cctaccaccg attctatgcc cccacttctg agacaagcaa 2641 ttgagctgtt cgaccggcag ggagccgaac ctgccttcct tttcggcctg gaactaatca 2701 tatgtggcct ggagaaacag ctaaagtgcg aaagcggcgg gccggccgac gcccttgacg 2761 attttgactt agacatgctc ccagccgatg cccttgacga ctttgacctt gatatgctgc 2821 ctgctgacgc tcttgacgat tttgaccttg acatgctccc cgggtaaatg catgaattcg 2881 atctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga 2941 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc 3001 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc 3061 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt 3121 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa 3181 gaaccagctg gggctcgaat caagctatca agtgccacct gacgtctccc tatcagtgat 3241 agagaagtcg acacgtctcg agctccctat cagtgataga gaaggtacgt ctagaacgtc 3301 tccctatcag tgatagagaa gtcgacacgt ctcgagctcc ctatcagtga tagagaaggt 3361 acgtctagaa cgtctcccta tcagtgatag agaagtcgac acgtctcgag ctccctatca 3421 gtgatagaga aggtaccccc tatataagca gagagatctg ttcaaatttg aactgactaa 3481 gcggctcccg ccagattttg gcaagattac taagcaggaa gtcaaggact tttttgcttg 3541 ggcaaaggtc aatcaggtgc cggtgactca cgagtttaaa gttcccaggg aattggcggg 3601 aactaaaggg gcggagaaat ctctaaaacg cccactgggt gacgtcacca atactagcta 3661 taaaagtctg gagaagcggg ccaggctctc atttgttccc gagacgcctc gcagttcaga 3721 cgtgactgtt gatcccgctc ctctgcgacc gctagcttcg atcaactacg cagacaggta 3781 ccaaaacaag tgttctcgtc acgtgggcat taatctgatt ctgtttccct gcagacaatg 3841 cgagagaatg aatcagaact caaatatctg cttcactcac ggacagaaag actgtttaga 3901 gtgctttccc gtgtcagaat ctcaacccgt ttctgtcgtc aaaaaggcgt atcagaaact 3961 gtgctacatt catcatatca tgggaaaggt gccagacgct tgcactgcct gcgatctggt 4021 caatgtggat ttggatgact gcatctttga acaataaatg acttaagcca ggtatggctg 4081 ccgatggtta tcttccagat tggctcgagg acaaccttag tgaaggaatt cgcgagtggt 4141 gggctttgaa acctggagcc cctcaaccca aggcaaatca acaacatcaa gacaacgcta 4201 gaggtcttgt gcttccgggt tacaaatacc ttggacccgg caacggactc gacaaggggg 4261 agccggtcaa cgcagcagac gcggcggccc tcgagcacga caaagcctac gaccagcagc 4321 tcaaggccgg agacaacccg tacctcaagt acaaccacgc cgacgccgag ttccaggagc 4381 ggctcaaaga agatacgtct tttgggggca acctcgggcg agcagtcttc caggccaaaa 4441 agaggcttct tgaacctctt ggtctggttg aggaagcggc taagacggct cctggaaaga 4501 agaggcctgt agagcagtct cctcaggaac cggactcctc cgcgggtatt ggcaaatcgg 4561 gtgcacagcc cgctaaaaag agactcaatt tcggtcagac tggcgacaca gagtcagtcc 4621 cagaccctca accaatcgga gaacctcccg cagccccctc aggtgtggga tctcttacaa 4681 tggcttcagg tggtggcgca ccagtggcag acaataacga aggtgccgat ggagtgggta 4741 gttcctcggg aaattggcat tgcgattccc aatggctggg ggacagagtc atcaccacca 4801 gcacccgaac ctgggccctg cccacctaca acaatcacct ctacaagcaa atctccaaca 4861 gcacatctgg aggatcttca aatgacaacg cctacttcgg ctacagcacc ccctgggggt 4921 attttgactt caacagattc cactgccact tctcaccacg tgactggcag cgactcatca 4981 acaacaactg gggattccgg cctaagcgac tcaacttcaa gctctttaac attcaggtca 5041 aagaggttac ggacaacaat ggagtcaaga ccatcgccaa taaccttacc agcacggtcc 5101 aggtcttcac ggactcagac tatcagctcc cgtacgtgct cgggtcggct cacgagggct 5161 gcctcccgcc gttcccagcg gacgttttca tgattcctca gtacgggtat ctgacgctta 5221 atgatggaag ccaggccgtg ggtcgttcgt ccttttactg cctggaatat ttcccgtcgc 5281 aaatgctaag aacgggtaac aacttccagt tcagctacga gtttgagaac gtacctttcc 5341 atagcagcta cgctcacagc caaagcctgg accgactaat gaatccactc atcgaccaat 5401 acttgtacta tctctctaga actattaacg gttctggaca gaatcaacaa acgctaaaat 5461 tcagtgtggc cggacccagc aacatggctg tccagggaag aaactacata cctggaccca 5521 gctaccgaca acaacgtgtc tcaaccactg tgactcaaaa caacaacagc gaatttgctt 5581 ggcctggagc ttcttcttgg gctctcaatg gacgtaatag cttgatgaat cctggacctg 5641 ctatggcctc tcacaaagaa ggagaggacc gtttctttcc tttgtctgga tctttaattt 5701 ttggcaaaca aggtactggc agagacaacg tggatgcgga caaagtcatg ataaccaacg 5761 aagaagaaat taaaactact aacccggtag caacggagtc ctatggacaa gtggccacaa 5821 accaccagag tgcccaaacc accctcaaac ccttcctcgc acaggcgcag accggttggg 5881 ttcaaaacca aggaatactt ccgggtatgg tttggcagga cagagatgtg tacctgcaag 5941 gacccatttg ggccaaaatt cctcacacgg acggcaactt tcacccttct ccgctgatgg 6001 gagggtttgg aatgaagcac ccgcctcctc agatcctcat caaaaacaca cctgtacctg 6061 cggatcctcc aacggccttc aacaaggaca agctgaactc tttcatcacc cagtattcta 6121 ctggtcaagt cagcgtggag atcgagtggg agctgcagaa ggaaaacagc aagcgctgga 6181 acccggagat ccagtacact tccaactatt acaagtctaa taatgttgaa tttgctgtta 6241 atactgaagg tgtatatagt gaaccccgcc ccattggcac cagatacctg actcgtaatc 6301 tgtaagtcga cttgcttgtt aatcaataaa ccgtttaatt cgtttcagtt gaactttggt 6361 ctctgcgaag ggcaattcgt ttaaacctgc aggactagag gtcctgtatt agaggtcacg 6421 tgagtgtttt gcgacatttt gcgacaccat gtggtcacgc tgggtattta agcccgagtg 6481 agcacgcagg gtctccattt tgaagcggga ggtttgaacg cgcagccgcc aagccgaatt 6541 ctgcagatat cacatgtcct aggaactatc gatccatcac actggcggcc gctcgactag 6601 agcggccgcc accgcggtgg agctccagct tttgcggacc gaatcggaaa gaacatgtga 6661 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 6721 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 6781 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 6841 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 6901 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 6961 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 7021 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 7081 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 7141 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 7201 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 7261 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 7321 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 7381 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 7441 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 7501 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 7561 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 7621 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 7681 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 7741 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 7801 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 7861 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 7921 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 7981 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 8041 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 8101 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 8161 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 8221 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 8281 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 8341 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 8401 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 8461 cctgacgtc //