LOCUS Exported 6224 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses the red fluorescent high-affinity DA sensor GRAB_rDA1h in neurons. ACCESSION . VERSION . KEYWORDS pAAV-hsyn-GRAB_rDA1h SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6224) AUTHORS Sun F, Zhou J, Dai B, Qian T, Zeng J, Li X, Zhuo Y, Zhang Y, Wang Y, Qian C, Tan K, Feng J, Dong H, Lin D, Cui G, Li Y. TITLE Next-generation GRAB sensors for monitoring dopaminergic activity in vivo. JOURNAL Nature Methods (2020) REFERENCE 2 (bases 1 to 6224) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6224 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" regulatory 581..590 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" sig_peptide 587..649 /product="leader sequence from mouse immunoglobulin kappa light chain" /label=Ig-kappa leader protein_bind 650..674 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" primer_bind 2044..2063 /label=mCherry-F /note="mCherry, forward primer" primer_bind complement(2404..2424) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2822..2833) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 2971..3447 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3170..3189) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3487..3627 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3487..3616 /label=AAV2 ITR rep_origin 3702..4157 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3789..3808) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3999..4020 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4174..4193) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4293..4315 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4353..4371) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4439..4543 /gene="bla" /label=AmpR promoter CDS 4544..5404 /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 complement(4762..4781) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5575..6163 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6064..6083 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt 61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 121 aggggttcct gcggccgcac gcgtatgagg cggggtgggg gtgcctacct gacgaccgac 181 cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag 241 agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg 301 acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg 361 ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc 421 gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc 481 gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga 541 ggagtcgtgt cgtgcctgag agcgcagtcg agaaaccggt gccaccatgg agacagacac 601 actcctgcta tgggtactgc tgctctgggt tccaggttcc actggtgaca caagtttgta 661 caaaaaagtt ggcaccaccg gtgatccact gaatctgtcc tggtatgatg atgatctgga 721 gaggcagaac tggagccggc ccttcaacgg gtcagacggg aaggcggaca gaccccacta 781 caactactat gccacactgc tcaccctgct catcgctgtc atcgtcttcg gcaacgtgct 841 ggtgtgcatg gctgtgtccc gcgagaaggc gctgcagacc accaccaact acctgatcgt 901 cagcctcgca gtggccgacc tcctcgtcgc cacactggtc atgccctggg ttgtctacct 961 ggaggtggta ggtgagtgga aattcagcag gattcactgt gacatcttcg tcactctgga 1021 cgtcatgatg tgcacggcga gcatcctgaa cttgtgtgcc atcagcatcg acaggtacac 1081 agctgtggcc atgcccatgc tgtacaatac gcgctacagc tccaagcgcc gggtcaccgt 1141 catgatctcc atcgtctggg tcctgtcctt caccatctcc tgcccactcc tcttcggact 1201 caataacgca gaccagaacg agtgcatcat tgccaacccg gccttcgtgg tctactcctc 1261 catcgtctcc ttctacgtgc ccttcattgt catgctgctg gtctacatca agatctacat 1321 tgtcctccgc agacgccgca agcgagtccc cgtgtgttcc gaggttatgt accccgagga 1381 cggcgccctg aagagcgaga tcaagaaggg gctgaggctg aaggacggcg gccactacgc 1441 cgccgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacat 1501 cgtcgacatc aagttggaca tcgtgtccca caacgaggac tacaccatcg tggaacagtg 1561 cgaacgcgcc gaggcccgcc actccaccgg cggcatggac gagctgtaca agggaggtac 1621 aggcgggagt ctggtgagca agggcgagga ggataacatg gccatcatca aggagttcat 1681 gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga 1741 gggcgagggc cgcccctacg aggcctttca gaccgctaag ctgaaggtga ccaagggtgg 1801 ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta 1861 cattaagcac ccagccgaca tccccgacta cttcaagctg tccttccccg agggcttcag 1921 gtgggagcgc gtgatgaact tcgaggacgg cggcattatt cacgttaacc aggactcctc 1981 cctgcaggac ggctacttca tctacaaggt gaagctgcgc ggcaccaact tcccccccga 2041 cggccccgta atgcagaaga agaccatggg ctgggagctg acggtccagc agctcgagaa 2101 gaaagccact cagatgctcg ccattgttct cggcgtgttc atcatctgct ggctgccctt 2161 cttcatcaca cacatcctga acatacactg tgactgcaac atcccgcctg tcctgtacag 2221 cgccttcacg tggctgggct atgtcaacag cgccgtgaac cccatcatct acaccacctt 2281 caacattgag ttccgcaagg ccttcctgaa gatcctccac tgctgagaat tcgatatcaa 2341 gcttatcgat catcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa 2401 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat 2461 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta 2521 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc 2581 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt 2641 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg 2701 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc 2761 ttggctgctc gcctatgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc 2821 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct 2881 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca 2941 tcgataccga gcgctgctcg agagatctac gggtggcatc cctgtgaccc ctccccagtg 3001 cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta 3061 agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg 3121 gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa 3181 ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa 3241 gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc 3301 agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc 3361 aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg 3421 tgaaccactg ctcccttccc tgtccttctg attttgtagg taaccacgtg cggaccgagc 3481 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca 3541 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga 3601 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc 3661 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc 3721 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt 3781 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg 3841 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga 3901 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt 3961 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg 4021 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc 4081 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat 4141 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt 4201 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc 4261 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc 4321 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt 4381 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg 4441 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca 4501 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt 4561 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga 4621 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga 4681 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat 4741 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca 4801 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt 4861 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac 4921 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct 4981 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga 5041 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac 5101 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat 5161 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg 5221 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc 5281 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc 5341 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg 5401 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta 5461 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg 5521 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga 5581 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt 5641 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag 5701 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa 5761 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag 5821 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca 5881 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac 5941 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa 6001 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc 6061 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg 6121 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc 6181 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt //