LOCUS       Exported                7193 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV expression of CaMKII-driven chimeric opsin variant C1V1 
            (E122T/E162T) for fast and potent optical excitation at red-shifted 
            wavelengths..
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-CaMKIIa-C1V1 (t/t)-TS-EYFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7193)
  AUTHORS   Yizhar O, Fenno LE, Prigge M, Schneider F, Davidson TJ, O'Shea DJ, 
            Sohal VS, Goshen I, Finkelstein J, Paz JT, Stehfest K, Fudim R, 
            Ramakrishnan C, Huguenard JR, Hegemann P, Deisseroth K
  TITLE     Neocortical excitation/inhibition balance in information processing 
            and social dysfunction.
  JOURNAL   Nature. 2011 Jul 27;477(7363):171-8. doi: 10.1038/nature10360.
  PUBMED    21796121
REFERENCE   2  (bases 1 to 7193)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7193
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   55..184
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        203..1491
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     CDS             2633..3349
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2675..2696)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2936..2955)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3283..3304
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3374..3962
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3427..3447)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3845..3856)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3994..4470
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4193..4212)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4510..4650
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4510..4639
                     /label=AAV2 ITR
     rep_origin      4725..5180
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4812..4831)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5022..5043
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5197..5216)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5316..5338
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5376..5394)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5462..5566
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5567..6427
                     /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(5785..5804)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6598..7186
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7087..7106
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtcctgca
       61 ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc gtcgggcgac ctttggtcgc
      121 ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt
      181 tcctgcggcc gcacgcgttt aacattatgg ccttaggtca cttcatctcc atggggttct
      241 tcttctgatt ttctagaaaa tgagatgggg gtgcagagag cttcctcagt gacctgccca
      301 gggtcacatc agaaatgtca gagctagaac ttgaactcag attactaatc ttaaattcca
      361 tgccttgggg gcatgcaagt acgatataca gaaggagtga actcattagg gcagatgacc
      421 aatgagttta ggaaagaaga gtccagggca gggtacatct acaccacccg cccagccctg
      481 ggtgagtcca gccacgttca cctcattata gttgcctctc tccagtccta ccttgacggg
      541 aagcacaagc agaaactggg acaggagccc caggagacca aatcttcatg gtccctctgg
      601 gaggatgggt ggggagagct gtggcagagg cctcaggagg ggccctgctg ctcagtggtg
      661 acagataggg gtgagaaagc agacagagtc attccgtcag cattctgggt ctgtttggta
      721 cttcttctca cgctaaggtg gcggtgtgat atgcacaatg gctaaaaagc agggagagct
      781 ggaaagaaac aaggacagag acagaggcca agtcaaccag accaattccc agaggaagca
      841 aagaaaccat tacagagact acaaggggga agggaaggag agatgaatta gcttcccctg
      901 taaaccttag aacccagctg ttgccagggc aacggggcaa tacctgtctc ttcagaggag
      961 atgaagttgc cagggtaact acatcctgtc tttctcaagg accatcccag aatgtggcac
     1021 ccactagccg ttaccatagc aactgcctct ttgccccact taatcccatc ccgtctgtta
     1081 aaagggccct atagttggag gtgggggagg taggaagagc gatgatcact tgtggactaa
     1141 gtttgttcgc atccccttct ccaaccccct cagtacatca ccctggggga acagggtcca
     1201 cttgctcctg ggcccacaca gtcctgcagt attgtgtata taaggccagg gcaaagagga
     1261 gcaggtttta aagtgaaagg caggcaggtg ttggggaggc agttaccggg gcaacgggaa
     1321 cagggcgttt cggaggtggt tgccatgggg acctggatgc tgacgaaggc tcgcgaggct
     1381 gtgagcagcc acagtgccct gctcagaagc cccaagctcg tcagtcaagc cggttctccg
     1441 tttgcactca ggagcacggg caggcgagtg gcccctagtt ctgggggcag ctctagagcg
     1501 gtaccggatc cccgggtacc ggtaggccac catgtcgcgg aggccatggc ttcttgccct
     1561 agcgctggca gtggcgctgg cggccggcag cgcaggagcc tcgactggca gtgacgcgac
     1621 ggtgccggtc gcgactcagg atggccccga ctacgttttc caccgtgccc acgagcgcat
     1681 gctcttccaa acctcataca ctcttgagaa caatggttct gttatttgca tcccgaacaa
     1741 cggccagtgc ttctgcttgg cttggcttaa atccaacgga acaaatgccg agaagttggc
     1801 tgccaacatt ctgcagtgga ttacttttgc gctttcagcg ctctgcctga tgttctacgg
     1861 ctaccagacc tggaagtcta cttgcggctg ggagaccatt tacgtggcca cgatcgagat
     1921 gatcaagttc atcatcgagt atttccatga gtttgacgaa cctgcggtga tctactcatc
     1981 caacggcaac aagaccgtgt ggcttcgtta cgcgacctgg ctgctcacct gcccggtcct
     2041 tctgatccat ctgagcaacc tcacaggcct gaaggacgat tatagcaaaa ggactatggg
     2101 cctgttggtt tctgatgtgg gatgcatcgt gtggggcgca accagcgcca tgtgtacggg
     2161 gtggacgaag atcctgttct tcctcatctc attgagctat ggtatgtata cctattttca
     2221 tgctgctaaa gtttatatcg aagcattcca cacagttcca aaagggattt gtcgagaact
     2281 ggtccgagtg atggcctgga cattctttgt ggcttgggga atgtttccag tcctgtttct
     2341 gctgggcacg gaaggattcg gtcatatcag cccttatgga tctgccattg ggcactccat
     2401 cctcgacctg attgcaaaga acatgtgggg tgtgctgggg aattacctgc gcgtcaaaat
     2461 ccacgagcac atcctgttgt atggcgacat cagaaagaag cagaaaatta cgatcgccgg
     2521 ccaagagatg gaggttgaga cactggtggc tgaagaggag gacgcggccg ccaagagcag
     2581 gatcaccagc gagggcgagt acatccccct ggaccagatc gacatcaacg tggtgagcaa
     2641 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     2701 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     2761 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     2821 cttcggctac ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
     2881 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     2941 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     3001 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     3061 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     3121 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     3181 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta
     3241 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     3301 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag aattcgatat
     3361 caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     3421 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     3481 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     3541 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     3601 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg gaactttcgc
     3661 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     3721 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     3781 tccttggctg ctcgcctatg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     3841 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     3901 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     3961 gcatcgatac cgagcgctgc tcgagagatc tacgggtggc atccctgtga cccctcccca
     4021 gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa
     4081 ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg
     4141 gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg
     4201 gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt
     4261 caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg
     4321 ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc
     4381 tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag
     4441 gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg
     4501 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     4561 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     4621 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
     4681 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     4741 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4801 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4861 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4921 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4981 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     5041 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     5101 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     5161 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     5221 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     5281 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     5341 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     5401 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     5461 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     5521 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     5581 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5641 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5701 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5761 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     5821 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5881 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5941 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     6001 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     6061 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     6121 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     6181 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     6241 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     6301 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     6361 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     6421 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     6481 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6541 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6601 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6661 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6721 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6781 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6841 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6901 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6961 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     7021 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     7081 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     7141 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg cca
//
