LOCUS       Exported                6459 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Optogenetics.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-hSyn-ChRmine-mScarlet-Kv2.1-WPRE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6459)
  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 6459)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6459
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      648..657
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1650..2342
                     /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(1780..1798)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2048..2067
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2586..3174
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2639..2659)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3057..3068)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3206..3682
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3405..3424)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3722..3862
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3722..3851
                     /label=AAV2 ITR
     rep_origin      3937..4392
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4024..4043)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4234..4255
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4409..4428)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4528..4550
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4588..4606)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4674..4778
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4779..5639
                     /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(4997..5016)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5810..6398
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6299..6318
                     /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 gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gatccccggg taccggtgcc accatggcac
      661 acgcaccagg caccgaccag atgttctacg tgggcacaat ggacggctgg tatctggata
      721 ccaagctgaa ctccgtggcc atcggcgccc actggtcttg ctttatcgtg ctgacaatca
      781 ccacattcta cctgggctat gagtcctgga catctcgggg ccctagcaag agaacctcct
      841 tttacgccgg ctatcaggag gagcagaacc tggccctgtt tgtgaatttc tttgccatgc
      901 tgagctactt cggcaagatc gtggccgaca cactgggcca caacttcggc gatgtgggcc
      961 ccttcatcat cggcttcggc aattacaggt atgccgacta catgctgacc tgcccaatgc
     1021 tggtgtacga tctgctgtat cagctgaggg ccccctatcg cgtgtcttgt agcgccatca
     1081 tctttgccat cctgatgtct ggcgtgctgg ccgagttcta cgcagagggc gaccctaggc
     1141 tgaggaatgg cgcctacgcc tggtatggct ttggctgttt ctggtttatc ttcgcctact
     1201 ctatcgtgat gagcatcgtg gccaagcagt atagccggct ggcccagctg gcccaggata
     1261 caggcgccga gcacagcctg cacgtgctga agttcgccgt gtttaccttc tccatgctgt
     1321 ggattctgtt tcccctggtg tgggccatct gccctagagg cttcggctgg atcgacgata
     1381 actggacaga ggtggcccac tgcgtgtgcg acatcgtggc caagtcctgt tacggctttg
     1441 ccctggcccg gttcagaaag acctatgatg aggagctgtt tcggctgctg gagcagctgg
     1501 gacacgacga ggatgagttc cagaagctgg agctggatat gaggctgagc agcaatggcg
     1561 agcgcctgcg gagactgtct gcggccgcca agagcaggat caccagcgag ggcgagtaca
     1621 tccccctgga ccagatcgac atcaacgtgg tgagcaaggg cgaggcagtg atcaaggagt
     1681 tcatgcggtt caaggtgcac atggagggct ccatgaacgg ccacgagttc gagatcgagg
     1741 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     1801 gtggccccct gcccttctcc tgggacatcc tgtcccctca gttcatgtac ggctccaggg
     1861 ccttcaccaa gcaccccgcc gacatccccg actactataa gcagtccttc cccgagggct
     1921 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgc cgtgaccgtg acccaggaca
     1981 cctccctgga ggacggcacc ctgatctaca aggtgaagct ccgcggcacc aacttccctc
     2041 ctgacggccc cgtaatgcag aagaagacaa tgggctggga agcgtccacc gagcggttgt
     2101 accccgagga cggcgtgctg aagggcgaca ttaagatggc cctgcgcctg aaggacggcg
     2161 ggcgctacct ggcggacttc aagaccacct acaaggccaa gaagcccgtg cagatgcccg
     2221 gcgcctacaa cgtggaccgc aagttggaca tcacctccca caacgaggac tacaccgtgg
     2281 tggaacagta cgaacgctcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     2341 agcagtccca gcccatcctc aacaccaagg agatggcccc gcagagcaag cctccagagg
     2401 agctggagat gagcagcatg cccagccccg tggcccctct gcccgcacgc acggagggcg
     2461 tcatcgacat gcggagcatg tccagcattg acagcttcat cagctgtgcc acggacttcc
     2521 ctgaagccac cagattcttc tgctacgaga acgaggtgta agaattcgat atcaagctta
     2581 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     2641 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     2701 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     2761 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     2821 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     2881 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     2941 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     3001 tgctcgccta tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     3061 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     3121 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     3181 accgagcgct gctcgagaga tctacgggtg gcatccctgt gacccctccc cagtgcctct
     3241 cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg
     3301 catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg
     3361 gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag
     3421 ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat
     3481 tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta
     3541 atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc
     3601 ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac
     3661 cactgctccc ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac cgagcggccg
     3721 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     3781 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     3841 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     3901 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     3961 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
     4021 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4081 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4141 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4201 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4261 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct
     4321 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     4381 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     4441 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     4501 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     4561 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     4621 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4681 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4741 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     4801 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     4861 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     4921 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     4981 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5041 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5101 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5161 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5221 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5281 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5341 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5401 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5461 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5521 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5581 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     5641 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     5701 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     5761 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5821 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5881 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5941 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6001 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6061 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6121 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6181 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6241 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6301 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6361 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6421 tacggttcct ggccttttgc tggccttttg ctcacatgt
//
