LOCUS Exported 8543 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION non-standard AAV2 rep-AAV-PHP.B4 cap plasmid with AAV cap expression controlled by a tTA-TRE amplification system. ACCESSION . VERSION . KEYWORDS pUCmini-iCAP-PHP.B4 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8543) 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 8543) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8543 /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" protein_bind 3449..3467 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 3486..3504 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" CDS 6221..6232 /codon_start=1 /product="WELQut protease recognition and cleavage site" /label=WELQut site /translation="WELQ" rep_origin complement(6788..7376) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6868..6887) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7547..8407) /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 8170..8189 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8408..8512) /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 aggtacgtct agaacgtctc cctatcagtg atagagaagt cgacacgtct 3481 cgagctccct atcagtgata gagaaggtac cccctatata agcagagaga tctgttcaaa 3541 tttgaactga ctaagcggct cccgccagat tttggcaaga ttactaagca ggaagtcaag 3601 gacttttttg cttgggcaaa ggtcaatcag gtgccggtga ctcacgagtt taaagttccc 3661 agggaattgg cgggaactaa aggggcggag aaatctctaa aacgcccact gggtgacgtc 3721 accaatacta gctataaaag tctggagaag cgggccaggc tctcatttgt tcccgagacg 3781 cctcgcagtt cagacgtgac tgttgatccc gctcctctgc gaccgctagc ttcgatcaac 3841 tacgcagaca ggtaccaaaa caagtgttct cgtcacgtgg gcattaatct gattctgttt 3901 ccctgcagac aatgcgagag aatgaatcag aactcaaata tctgcttcac tcacggacag 3961 aaagactgtt tagagtgctt tcccgtgtca gaatctcaac ccgtttctgt cgtcaaaaag 4021 gcgtatcaga aactgtgcta cattcatcat atcatgggaa aggtgccaga cgcttgcact 4081 gcctgcgatc tggtcaatgt ggatttggat gactgcatct ttgaacaata aatgacttaa 4141 gccaggtatg gctgccgatg gttatcttcc agattggctc gaggacaacc ttagtgaagg 4201 aattcgcgag tggtgggctt tgaaacctgg agcccctcaa cccaaggcaa atcaacaaca 4261 tcaagacaac gctagaggtc ttgtgcttcc gggttacaaa taccttggac ccggcaacgg 4321 actcgacaag ggggagccgg tcaacgcagc agacgcggcg gccctcgagc acgacaaagc 4381 ctacgaccag cagctcaagg ccggagacaa cccgtacctc aagtacaacc acgccgacgc 4441 cgagttccag gagcggctca aagaagatac gtcttttggg ggcaacctcg ggcgagcagt 4501 cttccaggcc aaaaagaggc ttcttgaacc tcttggtctg gttgaggaag cggctaagac 4561 ggctcctgga aagaagaggc ctgtagagca gtctcctcag gaaccggact cctccgcggg 4621 tattggcaaa tcgggtgcac agcccgctaa aaagagactc aatttcggtc agactggcga 4681 cacagagtca gtcccagacc ctcaaccaat cggagaacct cccgcagccc cctcaggtgt 4741 gggatctctt acaatggctt caggtggtgg cgcaccagtg gcagacaata acgaaggtgc 4801 cgatggagtg ggtagttcct cgggaaattg gcattgcgat tcccaatggc tgggggacag 4861 agtcatcacc accagcaccc gaacctgggc cctgcccacc tacaacaatc acctctacaa 4921 gcaaatctcc aacagcacat ctggaggatc ttcaaatgac aacgcctact tcggctacag 4981 caccccctgg gggtattttg acttcaacag attccactgc cacttctcac cacgtgactg 5041 gcagcgactc atcaacaaca actggggatt ccggcctaag cgactcaact tcaagctctt 5101 taacattcag gtcaaagagg ttacggacaa caatggagtc aagaccatcg ccaataacct 5161 taccagcacg gtccaggtct tcacggactc agactatcag ctcccgtacg tgctcgggtc 5221 ggctcacgag ggctgcctcc cgccgttccc agcggacgtt ttcatgattc ctcagtacgg 5281 gtatctgacg cttaatgatg gaagccaggc cgtgggtcgt tcgtcctttt actgcctgga 5341 atatttcccg tcgcaaatgc taagaacggg taacaacttc cagttcagct acgagtttga 5401 gaacgtacct ttccatagca gctacgctca cagccaaagc ctggaccgac taatgaatcc 5461 actcatcgac caatacttgt actatctctc tagaactatt aacggttctg gacagaatca 5521 acaaacgcta aaattcagtg tggccggacc cagcaacatg gctgtccagg gaagaaacta 5581 catacctgga cccagctacc gacaacaacg tgtctcaacc actgtgactc aaaacaacaa 5641 cagcgaattt gcttggcctg gagcttcttc ttgggctctc aatggacgta atagcttgat 5701 gaatcctgga cctgctatgg cctctcacaa agaaggagag gaccgtttct ttcctttgtc 5761 tggatcttta atttttggca aacaaggtac tggcagagac aacgtggatg cggacaaagt 5821 catgataacc aacgaagaag aaattaaaac tactaacccg gtagcaacgg agtcctatgg 5881 acaagtggcc acaaaccacc agagtgccca aacgttgcag attcctttta aggcacaggc 5941 gcagaccggt tgggttcaaa accaaggaat acttccgggt atggtttggc aggacagaga 6001 tgtgtacctg caaggaccca tttgggccaa aattcctcac acggacggca actttcaccc 6061 ttctccgctg atgggagggt ttggaatgaa gcacccgcct cctcagatcc tcatcaaaaa 6121 cacacctgta cctgcggatc ctccaacggc cttcaacaag gacaagctga actctttcat 6181 cacccagtat tctactggtc aagtcagcgt ggagatcgag tgggagctgc agaaggaaaa 6241 cagcaagcgc tggaacccgg agatccagta cacttccaac tattacaagt ctaataatgt 6301 tgaatttgct gttaatactg aaggtgtata tagtgaaccc cgccccattg gcaccagata 6361 cctgactcgt aatctgtaag tcgacttgct tgttaatcaa taaaccgttt aattcgtttc 6421 agttgaactt tggtctctgc gaagggcaat tcgtttaaac ctgcaggact agaggtcctg 6481 tattagaggt cacgtgagtg ttttgcgaca ttttgcgaca ccatgtggtc acgctgggta 6541 tttaagcccg agtgagcacg cagggtctcc attttgaagc gggaggtttg aacgcgcagc 6601 cgccaagccg aattctgcag atatcacatg tcctaggaac tatcgatcca tcacactggc 6661 ggccgctcga ctagagcggc cgccaccgcg gtggagctcc agcttttgcg gaccgaatcg 6721 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 6781 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 6841 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 6901 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 6961 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 7021 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 7081 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 7141 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 7201 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc 7261 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 7321 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 7381 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 7441 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 7501 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa 7561 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc 7621 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga 7681 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa 7741 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 7801 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 7861 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 7921 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg 7981 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 8041 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 8101 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 8161 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 8221 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 8281 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 8341 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 8401 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 8461 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 8521 cccgaaaagt gccacctgac gtc //