LOCUS       Exported                7089 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Optogenetics.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-CaMKIIa-ChRmine-mScarlet-WPRE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7089)
  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 7089)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7089
                     /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..3169
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGGA
                     VTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKM
                     ALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHS
                     TGGMDELYK"
     primer_bind     complement(2607..2625)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2875..2894
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3221..3809
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3274..3294)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3692..3703)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3841..4317
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4040..4059)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4357..4497
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4357..4486
                     /label=AAV2 ITR
     rep_origin      4572..5027
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4659..4678)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4869..4890
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5044..5063)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5163..5185
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5223..5241)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5309..5413
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5414..6274
                     /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(5632..5651)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6445..7033
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6934..6953
                     /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 ggcagtgatc aaggagttca tgcggttcaa
     2521 ggtgcacatg gagggctcca tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     2581 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     2641 cttctcctgg gacatcctgt cccctcagtt catgtacggc tccagggcct tcaccaagca
     2701 ccccgccgac atccccgact actataagca gtccttcccc gagggcttca agtgggagcg
     2761 cgtgatgaac ttcgaggacg gcggcgccgt gaccgtgacc caggacacct ccctggagga
     2821 cggcaccctg atctacaagg tgaagctccg cggcaccaac ttccctcctg acggccccgt
     2881 aatgcagaag aagacaatgg gctgggaagc gtccaccgag cggttgtacc ccgaggacgg
     2941 cgtgctgaag ggcgacatta agatggccct gcgcctgaag gacggcgggc gctacctggc
     3001 ggacttcaag accacctaca aggccaagaa gcccgtgcag atgcccggcg cctacaacgt
     3061 ggaccgcaag ttggacatca cctcccacaa cgaggactac accgtggtgg aacagtacga
     3121 acgctccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt tctgctacga
     3181 gaacgaggtg taatgagaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa
     3241 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3301 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3361 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     3421 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3481 ctgtcagctc ctttccggaa ctttcgcttt ccccctccct attgccacgg cggaactcat
     3541 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     3601 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg ccacctggat
     3661 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3721 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3781 tcggatctcc ctttgggccg cctccccgca tcgataccga gcgctgctcg agagatctac
     3841 gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca
     3901 gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc
     3961 ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca
     4021 acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg
     4081 ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg
     4141 ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg
     4201 ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct
     4261 tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg
     4321 attttgtagg taaccacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg
     4381 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     4441 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     4501 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     4561 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4621 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
     4681 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4741 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     4801 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4861 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
     4921 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
     4981 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     5041 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     5101 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     5161 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     5221 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     5281 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     5341 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     5401 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     5461 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     5521 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     5581 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     5641 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     5701 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     5761 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     5821 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     5881 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     5941 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     6001 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     6061 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     6121 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     6181 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     6241 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     6301 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     6361 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6421 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6481 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6541 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     6601 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6661 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6721 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6781 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6841 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6901 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6961 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     7021 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     7081 ttttgctca
//
