LOCUS       Exported                6373 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  ChR2-mCherry fusion expression in forebrain GABA-ergic interneurons 
            under the control of the mDlx enhancer element.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-mDlx-ChR2-mCherry-Fishell-3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6373)
  AUTHORS   Dimidschstein J, Chen Q, Tremblay R, Rogers SL, Saldi GA, Guo L, Xu 
            Q, Liu R, Lu C, Chu J, Avery MC, Rashid MS, Baek M, Jacob AL, Smith 
            GB, Wilson DE, Kosche G, Kruglikov I, Rusielewicz T, Kotak VC, 
            Mowery TM, Anderson SA, Callaway EM, Dasen JS, Fitzpatrick D, 
            Fossati V, Long MA, Noggle S, Reynolds JH, Sanes DH, Rudy B, Feng G,
            Fishell G
  TITLE     A viral strategy for targeting and manipulating interneurons across 
            vertebrate species.
  JOURNAL   Nat Neurosci. 2016 Oct 31. doi: 10.1038/nn.4430.
  PUBMED    27798629
REFERENCE   2  (bases 1 to 6373)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6373
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     intron          849..981
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     964..983
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1024..1033
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1030..1956
                     /codon_start=1
                     /product="H134R mutant of an N-terminal fragment of 
                     channelrhodopsin-2 from the alga Chlamydomonas reinhardtii 
                     (Nagel et al., 2005)"
                     /label=ChR2(H134R)
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIRLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     CDS             1969..2676
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2111..2129)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2379..2398
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2719..3307
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2772..2792)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3190..3201)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3476..3557
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3513..3532)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3567..3586
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3636..3776
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3636..3765
                     /label=AAV2 ITR
     rep_origin      3851..4306
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3938..3957)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4148..4169
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4323..4342)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4442..4464
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4502..4520)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4588..4692
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4693..5553
                     /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(4911..4930)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5724..6312
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6213..6232
                     /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 gcgtttaatt aagaattcta tacactcaca gtggtttggc
      181 atatatttgg tgaaattttt taaggaaaaa ttagtgttgg tttcgatata tggtagcttt
      241 ttctctaaca taatttgaat aattcagcaa agccctacta ccagctgtac ttctgcagcc
      301 tcttccattc tttccagcat tataattttg gttaattttc aattttaggt cctacgtctc
      361 tgcaatttgt gtatgaataa cagaataatt tccctctttt gtttcgcctt tcctgttcct
      421 gaatctaaat aaagatggct ttttagtatt aaaagtggaa gaaaattaca ggtaattatc
      481 tttgacggta aaaacgctgt aatcagcggg ctacatgaaa aattactcta attatggctg
      541 catttaagag aatggaaaaa aaccttcttg tggataaaaa ccttaaattg tccccaatgt
      601 ctgcttcaaa ttggatggca ctgcagctgg aggctttgtt cagaattgat cctggggagc
      661 tacgaaccca aagtttcaca gtaggaagct tgatatcgaa ttcctgcagc ccgggctggg
      721 cataaaagtc agggcagagc catctattgc ttacatttgc ttctagcctg caggtcgagg
      781 agcgcagcct tccagaagca gagcgcggcg ccttaagctg cagaagttgg tcgtgaggca
      841 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
      901 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
      961 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taagaaacta
     1021 gtagccacca tggactatgg cggcgctttg tctgccgtcg gacgcgaact tttgttcgtt
     1081 actaatcctg tggtggtgaa cgggtccgtc ctggtccctg aggatcaatg ttactgtgcc
     1141 ggatggattg aatctcgcgg cacgaacggc gctcagaccg cgtcaaatgt cctgcagtgg
     1201 cttgcagcag gattcagcat tttgctgctg atgttctatg cctaccaaac ctggaaatct
     1261 acatgcggct gggaggagat ctatgtgtgc gccattgaaa tggttaaggt gattctcgag
     1321 ttcttttttg agtttaagaa tccctctatg ctctaccttg ccacaggaca ccgggtgcag
     1381 tggctgcgct atgcagagtg gctgctcact tgtcctgtca tccttatccg cctgagcaac
     1441 ctcaccggcc tgagcaacga ctacagcagg agaaccatgg gactccttgt ctcagacatc
     1501 gggactatcg tgtggggggc taccagcgcc atggcaaccg gctatgttaa agtcatcttc
     1561 ttttgtcttg gattgtgcta tggcgcgaac acattttttc acgccgccaa agcatatatc
     1621 gagggttatc atactgtgcc aaagggtcgg tgccgccagg tcgtgaccgg catggcatgg
     1681 ctgtttttcg tgagctgggg tatgttccca attctcttca ttttggggcc cgaaggtttt
     1741 ggcgtcctga gcgtctatgg ctccaccgta ggtcacacga ttattgatct gatgagtaaa
     1801 aattgttggg ggttgttggg acactacctg cgcgtcctga tccacgagca catattgatt
     1861 cacggagata tccgcaaaac caccaaactg aacatcggcg gaacggagat cgaggtcgag
     1921 actctcgtcg aagacgaagc cgaggccgga gccgtgccag cggccgccgt gagcaagggc
     1981 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
     2041 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     2101 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
     2161 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
     2221 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     2281 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
     2341 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc
     2401 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
     2461 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
     2521 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
     2581 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
     2641 cactccaccg gcggcatgga cgagctgtac aagtaaaacg cgtgcccctg cagggaattc
     2701 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     2761 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     2821 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     2881 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     2941 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     3001 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3061 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     3121 tcctttcctt ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc
     3181 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     3241 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     3301 tccccgcatc gataccgagc gctgctcgag agatctgtga tagcggccat caagctggcc
     3361 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     3421 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3481 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3541 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcagctta
     3601 tcgataccgc atgcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc
     3661 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     3721 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     3781 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     3841 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3901 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
     3961 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     4021 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     4081 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4141 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
     4201 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
     4261 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     4321 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     4381 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     4441 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     4501 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     4561 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4621 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4681 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4741 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4801 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4861 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     4921 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     4981 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5041 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5101 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5161 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5221 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5281 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5341 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5401 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5461 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5521 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     5581 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5641 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5701 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5761 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5821 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5881 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5941 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6001 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6061 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6121 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6181 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6241 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6301 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6361 tttgctcaca tgt
//
