LOCUS       Exported                7623 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Anion channelrhodopsin GtACR1 fused to ER export signal and targeted
            to the neuronal soma and proximal dendrites under the control of 
            internal ribosome entry sequence in a viral vector.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-CAG-DIO-NLS-mRuby3-IRES-eGtACR1-ST
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7623)
  AUTHORS   Mardinly AR, Oldenburg IA, Pegard NC, Sridharan S, Lyall EH, Chesnov
            K, Brohawn SG, Waller L, Adesnik H
  TITLE     Precise multimodal optical control of neural ensemble activity.
  JOURNAL   Nat Neurosci. 2018 Apr 30. pii: 10.1038/s41593-018-0139-8. doi: 
            10.1038/s41593-018-0139-8.
  PUBMED    29713079
REFERENCE   2  (bases 1 to 7623)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7623
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     154..171
                     /label=L4440
                     /note="L4440 vector, forward primer"
     repeat_region   237..366
                     /label=AAV2 ITR
     polyA_signal    528..649
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(565..584)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     619..638
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     678..694
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     679..695
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    698..1286
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             804..815
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1213..1233
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    1311..1344
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    1395..1428
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     CDS             1441..1461
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1471..2181
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTD
                     IALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAV
                     AKYSNLGGGMDELYK"
     misc_feature    2216..2789
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2239..2791
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2383..2400)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2610..2629
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3717..3740
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    complement(3968..4001)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    complement(4052..4085)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     primer_bind     complement(4136..4155)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(4217..4239)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     repeat_region   5217..5346
                     /label=AAV2 ITR
     rep_origin      5418..5873
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5505..5524)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5715..5736
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5899..6003
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6004..6864
                     /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(6222..6241)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7035..7623
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7524..7543
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      121 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      181 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgc
      241 gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg acctttggtc
      301 gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg
      361 gttccttgta gttaatgatt aacccgccat gctacttatc tacgtagcca tgctctagga
      421 agatccttat cgattttacc acatttgtag aggttttact tgctttaaaa aacctcccac
      481 atctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
      541 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
      601 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgct
      661 cgaagcggcc gcccgggtcg aggtcgacgg tatcgatgcg gggaggcggc ccaaagggag
      721 atccgactcg tctgagggcg aaggcgaaga cgcggaagag gccgcagagc cggcagcagg
      781 ccgcgggaag gaaggtccgc tggattgagg gccgaaggga cgtagcagaa ggacgtcccg
      841 cgcagaatcc aggtggcaac acaggcgagc agccaaggaa aggacgatga tttccccgac
      901 aacaccacgg aattgtcagt gcccaacagc cgagcccctg tccagcagcg ggcaaggcag
      961 gcggcgatga gttccgccgt ggcaataggg agggggaaag cgaaagtccc ggaaaggagc
     1021 tgacaggtgg tggcaatgcc ccaaccagtg ggggttgcgt cagcaaacac agtgcacacc
     1081 acgccacgtt gcctgacaac gggccacaac tcctcataaa gagacagcaa ccaggattta
     1141 tacaaggagg agaaaatgaa agccatacgg gaagcaatag catgatacaa aggcattaaa
     1201 gcagcgtatc cacatagcgt aaaaggagca acatagttaa gaataccagt caatctttca
     1261 caaattttgt aatccagagg ttgattatcg ataagcttga tatcgaattc ataacttcgt
     1321 atagcataca ttatacgaag ttattctttg caccattcta aagaataaca gtgataattt
     1381 ctgggttaag gcaaataact tcgtatagga tactttatac gaagttatgc tagcatggca
     1441 cccaagaaga agaggaaggt ggcggccgct atggtgtcta agggcgaaga gctgatcaag
     1501 gaaaatatgc gtatgaaggt ggtcatggaa ggttcggtca acggccacca attcaaatgc
     1561 acaggtgaag gagaaggcag accgtacgag ggagtgcaaa ccatgaggat caaagtcatc
     1621 gagggaggac ccctgccatt tgcctttgac attcttgcca cgtcgttcat gtatggcagc
     1681 cgtaccttta tcaagtaccc ggccgacatc cctgatttct ttaaacagtc ctttcctgag
     1741 ggttttactt gggaaagagt tacgagatac gaagatggtg gagtcgtcac cgtcacgcag
     1801 gacaccagcc ttgaggatgg cgagctcgtc tacaacgtca aggtcagagg ggtaaacttt
     1861 ccctccaatg gtcccgtgat gcagaagaag accaagggtt gggagcctaa tacagagatg
     1921 atgtatccag cagatggtgg tctgagagga tacactgaca tcgcactgaa agttgatggt
     1981 ggtggccatc tgcactgcaa cttcgtgaca acttacaggt caaaaaagac cgtcgggaac
     2041 atcaagatgc ccggtgtcca tgccgttgat caccgcctgg aaaggatcga ggagagtgac
     2101 aatgaaacct acgtagtgca aagagaagtg gcagttgcca aatacagcaa ccttggtggt
     2161 ggcatggacg agctgtacaa gtgaattcct cgactagaat tgggcccggg atccgcccct
     2221 ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     2281 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     2341 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     2401 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
     2461 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     2521 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     2581 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     2641 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtacacat
     2701 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
     2761 tggttttcct ttgaaaaaca cgatgataat atggccacag cttgtccacc atgctcgagt
     2821 ccatgagcag catcacctgt gatcccgcca tctacggcga atggagcagg gagaaccagt
     2881 tctgcgtgga gaagagcctg atcaccctgg acggcatcaa gtacgtccag ctggtgatgg
     2941 ccgtcgtgag cgcctgtcag gtgttcttca tggtgaccag agcccccaag gtgccctggg
     3001 aagccatcta cctgcccacc accgagatga tcacctattc cctggccttc accggaaacg
     3061 gctacatcag agtggccaat ggcaagtacc tgccctgggc cagaatggcc agctggctgt
     3121 gcacctgccc tatcatgctg ggcctggtga gcaatatggc cctcgtgaag tacaagagca
     3181 tccccctcaa ccctatgatg atcgccgcct ccagcatctg caccgtgttc ggcatcaccg
     3241 cctccgtggt gctagacccg ctgcacgtgt ggctgtactg tttcatcagc agcatcttct
     3301 tcatcttcga gatggtggtg gccttcgcca ttttcgccat taccatccac gatttccaga
     3361 ccatcggctc ccccatgtcc ctgaaggtgg tggagaggct gaagctgatg aggatcgtgt
     3421 tctacgtgag ctggatggcc taccctatcc tgtggagctt ctccagcacc ggcgcctgca
     3481 tcatgagcga gaacaccagc agcgtgctgt acctgctggg cgacgctctg tgcaagaaca
     3541 cctacggcat cctgctgtgg gctacaacct ggggcctgct gaacggcaag tgggacaggg
     3601 attacgtgaa gggcaggaac gtggacggca ccctgatgcc tgagtacgag caggacctgg
     3661 agaagggcaa caccgagagg tacgaggacg ccagagccgg cgagaccgca ccggtagatt
     3721 acaaagacga tgacgataag cagtcccagc ccatcctcaa caccaaggag atggccccgc
     3781 agagcaagcc tccagaggag ctggagatga gcagcatgcc cagccccgtg gcccctctgc
     3841 ccgcacgcac ggagggcgtc atcgacatgc ggagcatgtc cagcattgac agcttcatca
     3901 gctgtgccac ggacttccct gaagccacca gattcttctg ctacgagaac gaggtgtgag
     3961 gcgcgccata acttcgtata atgtatgcta tacgaagtta ttaagaggtt tcatattgct
     4021 aatagcagct acaatccagc taccattctg cataacttcg tataaagtat cctatacgaa
     4081 gttattccgg agtcgactct agaggatcca attctttgcc aaaatgatga gacagcacaa
     4141 taaccagcac gttgcccagg agctgtagga aaaagaagaa ggcatgaaca tggttagcag
     4201 aggctctaga gccgccggtc acacgccaga agccgaaccc cgccctgccc cgtccccccc
     4261 gaaggcagcc gtccccccgc ggacagcccc gagccgctcc ccccgcacaa agggccctcc
     4321 cggagcccct caaggctttc acgcagccac agaaaagaaa caagccgtca ttaaaccaag
     4381 cgctaattac agcccggagg agaagggccg tcccgcccgc tcacctgtgg gagtaacgcg
     4441 gtcagtcaga gccggggcgg gcggcgcgag gcggcggcgg agcggggcac ggggcgaagg
     4501 cagcgcgcag cgactcccgc ccgccgcgcg cttcgctttt tatagggccg ccgccgccgc
     4561 cgcctcgcca taaaaggaaa ctttcggagc gcgccgctct gattggctgc cgccgcacct
     4621 ctccgcctcg ccccgccccg cccctcgccc cgccccgccc cgcctggcgc gcgccccccc
     4681 cccccccccc cccccccccc cccccccccc cccccccccc cccccccccc cccccccccc
     4741 cccccccccc cccccccccc cccccccccc cccnsccscc cccccccccc cccccccgcc
     4801 cccatcgctg cacaaaataa ttaaaaaata aataaataca aaattggggg tggggagggg
     4861 ggggagatgg ggagagtgaa gcagaacgtg gggctcacct cgaccatggt aatagcgatg
     4921 actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa
     4981 tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac
     5041 acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg
     5101 aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tggtacagat
     5161 cttcctagag catggctacg tagataagta gcatggcggg ttaatcatta actacaagga
     5221 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     5281 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     5341 gcgcagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     5401 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     5461 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     5521 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     5581 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     5641 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5701 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     5761 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     5821 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     5881 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5941 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     6001 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     6061 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     6121 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     6181 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     6241 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     6301 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     6361 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     6421 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     6481 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     6541 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     6601 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     6661 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     6721 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6781 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6841 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6901 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6961 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     7021 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     7081 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     7141 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     7201 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     7261 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     7321 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     7381 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     7441 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     7501 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     7561 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7621 aaa
//
