LOCUS Exported 7099 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Chloride Channel. ACCESSION . VERSION . KEYWORDS pAAV-CaMKIIa-SwiChRca-TS-EYFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7099) AUTHORS Berndt A, Lee SY, Ramakrishnan C, Deisseroth K TITLE Structure-guided transformation of channelrhodopsin into a light-activated chloride channel. JOURNAL Science. 2014 Apr 25;344(6182):420-4. doi: 10.1126/science.1252367. PUBMED 24763591 REFERENCE 2 (bases 1 to 7099) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7099 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..141 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 12..141 /label=AAV2 ITR promoter 160..1448 /label=CaMKII promoter /note="promoter for the mouse calcium/calmodulin-dependent kinase II alpha subunit" regulatory 1483..1492 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2485..3201 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFREDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(2527..2548) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2788..2807) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3135..3156 /label=EGFP-C /note="EGFP, forward primer" misc_feature 3226..3814 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3279..3299) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3697..3708) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3846..4322 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4045..4064) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4362..4502 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4362..4491 /label=AAV2 ITR rep_origin 4577..5032 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4664..4683) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4874..4895 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5049..5068) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5168..5190 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5228..5246) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5314..5418 /gene="bla" /label=AmpR promoter CDS 5419..6279 /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(5637..5656) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6450..7038 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6939..6958 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 121 actccatcac taggggttcc tgcggccgca cgcgtttaac attatggcct taggtcactt 181 catctccatg gggttcttct tctgattttc tagaaaatga gatgggggtg cagagagctt 241 cctcagtgac ctgcccaggg tcacatcaga aatgtcagag ctagaacttg aactcagatt 301 actaatctta aattccatgc cttgggggca tgcaagtacg atatacagaa ggagtgaact 361 cattagggca gatgaccaat gagtttagga aagaagagtc cagggcaggg tacatctaca 421 ccacccgccc agccctgggt gagtccagcc acgttcacct cattatagtt gcctctctcc 481 agtcctacct tgacgggaag cacaagcaga aactgggaca ggagccccag gagaccaaat 541 cttcatggtc cctctgggag gatgggtggg gagagctgtg gcagaggcct caggaggggc 601 cctgctgctc agtggtgaca gataggggtg agaaagcaga cagagtcatt ccgtcagcat 661 tctgggtctg tttggtactt cttctcacgc taaggtggcg gtgtgatatg cacaatggct 721 aaaaagcagg gagagctgga aagaaacaag gacagagaca gaggccaagt caaccagacc 781 aattcccaga ggaagcaaag aaaccattac agagactaca agggggaagg gaaggagaga 841 tgaattagct tcccctgtaa accttagaac ccagctgttg ccagggcaac ggggcaatac 901 ctgtctcttc agaggagatg aagttgccag ggtaactaca tcctgtcttt ctcaaggacc 961 atcccagaat gtggcaccca ctagccgtta ccatagcaac tgcctctttg ccccacttaa 1021 tcccatcccg tctgttaaaa gggccctata gttggaggtg ggggaggtag gaagagcgat 1081 gatcacttgt ggactaagtt tgttcgcatc cccttctcca accccctcag tacatcaccc 1141 tgggggaaca gggtccactt gctcctgggc ccacacagtc ctgcagtatt gtgtatataa 1201 ggccagggca aagaggagca ggttttaaag tgaaaggcag gcaggtgttg gggaggcagt 1261 taccggggca acgggaacag ggcgtttcgg aggtggttgc catggggacc tggatgctga 1321 cgaaggctcg cgaggctgtg agcagccaca gtgccctgct cagaagcccc aagctcgtca 1381 gtcaagccgg ttctccgttt gcactcagga gcacgggcag gcgagtggcc cctagttctg 1441 ggggcagctc tagagcggta ccggatcccc gggtaccggt aggccaccat ggactatggc 1501 ggcgctttgt ctgccgtcgg actcttccaa acctcataca ctcttgagaa caatggttct 1561 gttatttgca tcccgaacaa cggccagtgc ttctgcttgg cttggcttaa atccaacgga 1621 acaaatgccg agaagttggc tgccaacatt ctgcagtgga tttcttttgc gctttcagcg 1681 ctctgcctga tgttctacgg ctaccagacc tggaagtcta cttgcggctg ggaggagatt 1741 tacgtggcca cgatctcgat gatcaagttc atcatcgagt atttccatag ctttgacgaa 1801 cctgcggtga tctactcatc caacggcaac aagaccaagt ggcttcgtta cgcgagctgg 1861 ctgctgaccg cccctgtcat tcttatccgt ctgagcaacc ttacgggtct ggcgaacgac 1921 tataacaagc gtaccatggg tctgctggtg tcagatatcg gcacgatcgt gtggggcacc 1981 acggccgcgc tgtccaaggg atacgtccgt gtcattttct tcctgatggg cctgtgctac 2041 ggcatctaca cattcttcaa cgcagccaag gtctacattg aggcgtacca caccgtgccg 2101 aagggccggt gtcgccaggt ggtgactggc atggcttggc tcttcttcgt atcatggggt 2161 atgttcccca tcctgttcat cctcggcccc gagggcttcg gcgtcctgag caagtacggc 2221 tccaacgtcg gccacaccat cattgacctg atgtcgaagc agtgctgggg tctgctcggc 2281 cactacctgc gcgtgctgat ccacgagcat atcctcatcc acggcgacat tcgcaagacc 2341 accaaattga acattggtgg cactgagatt gaggtcgaga cgctggtgga ggacgaggcc 2401 gaggctggcg cggtcgcggc cgccaagagc aggatcacca gcgagggcga gtacatcccc 2461 ctggaccaga tcgacatcaa cgtggtgagc aagggcgagg agctgttcac cggggtggtg 2521 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag 2581 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag 2641 ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcctgca gtgcttcgcc 2701 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac 2761 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg 2821 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcagggag 2881 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc 2941 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag 3001 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc 3061 gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa agaccccaac 3121 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc 3181 atggacgagc tgtacaagta agaattcgat atcaagctta tcgataatca acctctggat 3241 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 3301 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 3361 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 3421 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 3481 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 3541 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 3601 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc 3661 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 3721 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 3781 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga 3841 tctacgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca 3901 ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg 3961 tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga 4021 agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat 4081 cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg 4141 agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga 4201 gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc 4261 caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc 4321 ttctgatttt gtaggtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga 4381 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc 4441 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca 4501 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg 4561 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt 4621 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc 4681 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct 4741 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat 4801 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc 4861 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcgggc 4921 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg 4981 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc 5041 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca 5101 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg 5161 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga 5221 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct 5281 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc 5341 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa 5401 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt 5461 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct 5521 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc 5581 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta 5641 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac 5701 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc 5761 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac 5821 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg 5881 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac 5941 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc 6001 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt 6061 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga 6121 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc 6181 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag 6241 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca 6301 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc 6361 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca 6421 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc 6481 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta 6541 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt 6601 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc 6661 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg 6721 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg 6781 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag 6841 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc 6901 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat 6961 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg 7021 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc 7081 tggccttttg ctcacatgt //