LOCUS       Exported                7302 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Optogenetics.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-CaMKIIa-ChRmine-eYFP-Kv2.1-WPRE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7302)
  AUTHORS   Marshel JH, Kim YS, Machado TA, Quirin S, Benson B, Kadmon J, Raja 
            C, Chibukhchyan A, Ramakrishnan C, Inoue M, Shane JC, McKnight DJ, 
            Yoshizawa S, Kato HE, Ganguli S, Deisseroth K
  TITLE     Cortical layer-specific critical dynamics triggering perception.
  JOURNAL   Science. 2019 Aug 9;365(6453). pii: science.aaw5202. doi: 
            10.1126/science.aaw5202. Epub 2019 Jul 18.
  PUBMED    31320556
REFERENCE   2  (bases 1 to 7302)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7302
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   6..135
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        154..1442
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1475..1484
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2477..3190
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2519..2540)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2780..2799)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3127..3148
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3434..4022
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3487..3507)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3905..3916)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4054..4530
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4253..4272)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4570..4710
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4570..4699
                     /label=AAV2 ITR
     rep_origin      4785..5240
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4872..4891)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5082..5103
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5257..5276)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5376..5398
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5436..5454)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5522..5626
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5627..6487
                     /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(5845..5864)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6658..7246
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7147..7166
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga
       61 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
      121 tcactagggg ttcctgcggc cgcacgcgtt taacattatg gccttaggtc acttcatctc
      181 catggggttc ttcttctgat tttctagaaa atgagatggg ggtgcagaga gcttcctcag
      241 tgacctgccc agggtcacat cagaaatgtc agagctagaa cttgaactca gattactaat
      301 cttaaattcc atgccttggg ggcatgcaag tacgatatac agaaggagtg aactcattag
      361 ggcagatgac caatgagttt aggaaagaag agtccagggc agggtacatc tacaccaccc
      421 gcccagccct gggtgagtcc agccacgttc acctcattat agttgcctct ctccagtcct
      481 accttgacgg gaagcacaag cagaaactgg gacaggagcc ccaggagacc aaatcttcat
      541 ggtccctctg ggaggatggg tggggagagc tgtggcagag gcctcaggag gggccctgct
      601 gctcagtggt gacagatagg ggtgagaaag cagacagagt cattccgtca gcattctggg
      661 tctgtttggt acttcttctc acgctaaggt ggcggtgtga tatgcacaat ggctaaaaag
      721 cagggagagc tggaaagaaa caaggacaga gacagaggcc aagtcaacca gaccaattcc
      781 cagaggaagc aaagaaacca ttacagagac tacaaggggg aagggaagga gagatgaatt
      841 agcttcccct gtaaacctta gaacccagct gttgccaggg caacggggca atacctgtct
      901 cttcagagga gatgaagttg ccagggtaac tacatcctgt ctttctcaag gaccatccca
      961 gaatgtggca cccactagcc gttaccatag caactgcctc tttgccccac ttaatcccat
     1021 cccgtctgtt aaaagggccc tatagttgga ggtgggggag gtaggaagag cgatgatcac
     1081 ttgtggacta agtttgttcg catccccttc tccaaccccc tcagtacatc accctggggg
     1141 aacagggtcc acttgctcct gggcccacac agtcctgcag tattgtgtat ataaggccag
     1201 ggcaaagagg agcaggtttt aaagtgaaag gcaggcaggt gttggggagg cagttaccgg
     1261 ggcaacggga acagggcgtt tcggaggtgg ttgccatggg gacctggatg ctgacgaagg
     1321 ctcgcgaggc tgtgagcagc cacagtgccc tgctcagaag ccccaagctc gtcagtcaag
     1381 ccggttctcc gtttgcactc aggagcacgg gcaggcgagt ggcccctagt tctgggggca
     1441 gctctagagc ggtaccggat ccccgggtac cggtgccacc atggcacacg caccaggcac
     1501 cgaccagatg ttctacgtgg gcacaatgga cggctggtat ctggatacca agctgaactc
     1561 cgtggccatc ggcgcccact ggtcttgctt tatcgtgctg acaatcacca cattctacct
     1621 gggctatgag tcctggacat ctcggggccc tagcaagaga acctcctttt acgccggcta
     1681 tcaggaggag cagaacctgg ccctgtttgt gaatttcttt gccatgctga gctacttcgg
     1741 caagatcgtg gccgacacac tgggccacaa cttcggcgat gtgggcccct tcatcatcgg
     1801 cttcggcaat tacaggtatg ccgactacat gctgacctgc ccaatgctgg tgtacgatct
     1861 gctgtatcag ctgagggccc cctatcgcgt gtcttgtagc gccatcatct ttgccatcct
     1921 gatgtctggc gtgctggccg agttctacgc agagggcgac cctaggctga ggaatggcgc
     1981 ctacgcctgg tatggctttg gctgtttctg gtttatcttc gcctactcta tcgtgatgag
     2041 catcgtggcc aagcagtata gccggctggc ccagctggcc caggatacag gcgccgagca
     2101 cagcctgcac gtgctgaagt tcgccgtgtt taccttctcc atgctgtgga ttctgtttcc
     2161 cctggtgtgg gccatctgcc ctagaggctt cggctggatc gacgataact ggacagaggt
     2221 ggcccactgc gtgtgcgaca tcgtggccaa gtcctgttac ggctttgccc tggcccggtt
     2281 cagaaagacc tatgatgagg agctgtttcg gctgctggag cagctgggac acgacgagga
     2341 tgagttccag aagctggagc tggatatgag gctgagcagc aatggcgagc gcctgcggag
     2401 actgtctgcg gccgccaaga gcaggatcac cagcgagggc gagtacatcc ccctggacca
     2461 gatcgacatc aacgtggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     2521 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     2581 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     2641 gccctggccc accctcgtga ccaccttcgg ctacggcctg cagtgcttcg cccgctaccc
     2701 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     2761 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     2821 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     2881 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     2941 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     3001 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     3061 gcccgacaac cactacctga gctaccagtc cgccctgagc aaagacccca acgagaagcg
     3121 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     3181 gctgtacaag cagtcccagc ccatcctcaa caccaaggag atggccccgc agagcaagcc
     3241 tccagaggag ctggagatga gcagcatgcc cagccccgtg gcccctctgc ccgcacgcac
     3301 ggagggcgtc atcgacatgc ggagcatgtc cagcattgac agcttcatca gctgtgccac
     3361 ggacttccct gaagccacca gattcttctg ctacgagaac gaggtgtaag aattcgatat
     3421 caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     3481 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     3541 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     3601 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     3661 cgcaaccccc actggttggg gaattgccac cacctgtcag ctcctttccg gaactttcgc
     3721 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     3781 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     3841 tccttggctg ctcgcctatg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     3901 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     3961 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     4021 gcatcgatac cgagcgctgc tcgagagatc tacgggtggc atccctgtga cccctcccca
     4081 gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa
     4141 ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg
     4201 gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg
     4261 gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt
     4321 caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg
     4381 ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc
     4441 tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag
     4501 gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg
     4561 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     4621 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     4681 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
     4741 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     4801 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4861 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4921 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4981 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     5041 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     5101 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     5161 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     5221 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     5281 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     5341 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     5401 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     5461 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     5521 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     5581 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     5641 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5701 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5761 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5821 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     5881 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5941 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     6001 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     6061 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     6121 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     6181 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     6241 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     6301 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     6361 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     6421 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     6481 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     6541 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6601 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6661 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6721 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6781 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6841 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6901 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6961 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     7021 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     7081 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     7141 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     7201 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     7261 ggccttttta cggttcctgg ccttttgctg gccttttgct ca
//
