LOCUS       Exported                7504 bp DNA     circular SYN 13-JUL-2021
DEFINITION  Cation channelrhodopsin ChrimsonR fused to mRuby2 fluorophore and 
            targeted to the neuronal soma and proximal dendrites in a viral 
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7504)
  AUTHORS   Pegard NC, Mardinly AR, Oldenburg IA, Sridharan S, Waller L, Adesnik
            H
  TITLE     Three-dimensional scanless holographic optogenetics with temporal 
            focusing (3D-SHOT).
  JOURNAL   Nat Commun. 2017 Oct 31;8(1):1228. doi: 10.1038/s41467-017-01031-3.
  PUBMED    29089483
REFERENCE   2  (bases 1 to 7504)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2021 from SnapGene 5.3.2
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2017-10-31"; volume: "8(1):1228. doi"; pages: " 
            10.1038/s41467-017-01031-3"
FEATURES             Location/Qualifiers
     source          1..7504
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   39..168
                     /label=AAV2 ITR
     promoter        250..697
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    735..768
                     /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."
     protein_bind    819..852
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     primer_bind     853..883
                     /label=X2053ST_Asc_R
     misc_feature    complement(861..1067)
                     /label=ST
     primer_bind     complement(1051..1081)
                     /label=X2052Chr_ST_Fm
     primer_bind     1060..1085
                     /label=X2051Chr_ST_Rm
     misc_feature    complement(1068..2129)
                     /label=ChrimsonR
     primer_bind     complement(2111..2146)
                     /label=X2050P2A_Chr_Fm
     primer_bind     2122..2152
                     /label=X2049P2A_ChR_Rm
     CDS             complement(2136..2192)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(2180..2213)
                     /label=X2048jGCa_P2_Fm
     primer_bind     2193..2229
                     /label=X2047jGCa_P2_Rm
     misc_feature    complement(2202..3473)
                     /label=jGCaMP8s
     primer_bind     2758..2777
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3017..3038
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3144..3165)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(3447..3464)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3453..3479)
                     /label=X2046jGCa_NheI_F
     protein_bind    complement(3480..3513)
                     /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."
     protein_bind    complement(3564..3597)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3622..4210
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3675..3695)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4093..4104)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4213..4229)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4214..4230)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    4259..4380
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4270..4289)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4324..4343
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4542..4671
                     /label=AAV2 ITR
     rep_origin      complement(5266..5854)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5346..5365)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6025..6885)
                     /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     6648..6667
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6886..6990)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7016..7471)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7153..7174)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct gcgcgctcgc tcgctcactg
       61 aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg
      121 agcgagcgcg cagagaggga gtggccaact ccatcactag gggttccttg tagttaatga
      181 ttaacccgcc atgctactta tctacgtagc catgctctag gaagatctct gcagagggcc
      241 ctgcgtatga gtgcaagtgg gttttaggac caggatgagg cggggtgggg gtgcctacct
      301 gacgaccgac cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc
      361 cctatcagag agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc
      421 agcaccgcgg acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac
      481 tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg
      541 tcgcgtccgc gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg
      601 gcacgggcgc gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt
      661 gggcagcgga ggagtcgtgt cgtgcctgag agcgcagtcg agaaggtacc ggatcctcta
      721 gagtcgactc cggaataact tcgtatagga tactttatac gaagttatgc agaatggtag
      781 ctggattgta gctgctatta gcaatatgaa acctcttaat aacttcgtat agcatacatt
      841 atacgaagtt atggcgcgcc ttagaatctg gtggcttcag ggaagtccgt ggcacagctg
      901 atgaagctgt caatgctgga catgctccgc atgtcgatga cgccctccgt gcgtgcgggc
      961 agaggggcca cggggctggg catgctgctc atctccagct cctctggagg cttgctctgc
     1021 ggggccatct ccttggtgtt gaggatgggc tgggactgct ctagggctac cggtgccgcc
     1081 actgtgtcct cgtcctcctc ctccacgaac tcttccactt ccacctcctc gcctccgatc
     1141 tccatcttgg tggtcttccg gatgtcgccg tggatcagga tgtgctcgtg gatcttgatc
     1201 ctcaggtggt gggccaggaa ggtccaaaac tccttggcga tgatgtcgca gatgctgtgg
     1261 ccgatgctgt tggcgtaagg gctcagcttc agcagtcctt ctggtcccac tgcccagagg
     1321 attgggtagc tgccccaaga ggcgaagtaa gcgtaggcca tcagcttcac gaccatgcgg
     1381 caatggcctt taggcacgct gtggttggct tccacgtagc acttggcggc ctggaagtac
     1441 atgtagccgc cgtagatgca agacacgata tacagcagcc acttgagcca atcggtagcc
     1501 agtcctgcgg ccatgccgaa cacgatcatt cccacgcaag acacgatcag gcccatggtc
     1561 cgcttgctgt agtcgttctt caggccgctc aggttgctca gtctgatcag gatcacgggg
     1621 caagacagca gccactcgaa gtagcgcagg caataggcgt ggttgccggt agacaggtac
     1681 actgtggcgg ggctgctgaa ctccttgaag atctccaggg tcacgaacag cacctcgacg
     1741 caacagacgt agacctcctc ccaaccgcaa gtggccttcc aggcgctgaa gccgtagaat
     1801 gtcagcaggg cgatggcgat gctgaaagca atccactggc agacctgggc gccgatcttt
     1861 tctcctggtg tgcctctgct gtgcagccaa gcctgcttct tctccactcc cacgaccagg
     1921 tagccgcctg tggcctccac gaagcaatat ccgtatccgg cgtcgctcag tccgtaagaa
     1981 gggtcacacc accactcggt gctgaagcac tcgcctgtag gtgtcattcc tccacagccc
     2041 atgtcctcgt ggtggtaggg gttaggccaa gggttgatgc ctccggcggc aaacagagat
     2101 ctggtggcgc tgctgatcag ctcagccata ctagtaggtc cagggttctc ctccacgtct
     2161 ccagcctgct tcagcaggct gaagttagta gctccgcttc ccttcgctgt catcatttgt
     2221 acaaactctt cgtagtttac ctgaccatcc ccatcgatgt ctgcttccct gatcatttca
     2281 tcaacctctt catctgttaa cttctctcca aggtttgtca tcacgtggcg aagctctgct
     2341 gcactgatgt agccattgcc atccttatca aacacaccga acgcttctct aatttcttct
     2401 tccgtgtccc tgtatttcat ttttcttgcc atcattgtca ggaactcagg gaagtcgatt
     2461 gtgccgtcac cgtcggcatc tacttcattg atcatgtcct gcagctctgc ttctgtgggg
     2521 ttgtgtccga gagaccgcat caccgtcccc agctccttgg ttgttattgt cccatccccg
     2581 tccttgtcaa atagggagaa agcctctttg tattctgcga tctgctcttc agtcagttgg
     2641 tccggcaggt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
     2701 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
     2761 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggatgtagcc ttcgggcatg
     2821 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg
     2881 ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
     2941 atgaacttca gggtcagctt gccgtaggtg gcatcgccct caccctcgcc ggacacgctg
     3001 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
     3061 agctcctcgc ccttgctcac catgctccct ccggtaccgc ccttgtacag ctcgtccatg
     3121 ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg
     3181 ttggggtctt tcgaaagttt ggactccacg ctcaggtagt ggttgtcggg cagcagcacg
     3241 gggccgtcgc cgatgggggt gttctgctgg tagtggtagg cgagctgcac gccgccgtcc
     3301 tcgatgttgt ggcggatgtg gaagttcgcc ttgatgccgt tcttctgctt gtcggccttg
     3361 atatagacgt tgatcttcag gcccatcagc atggcgatga tcttcacggc ggtggccacc
     3421 tccttgaagg tcttcttgcg acgcgtgtga tgatggtgat gatgcatggt ggcgctagca
     3481 taacttcgta taaagtatcc tatacgaagt tatttgcctt aacccagaaa ttatcactgt
     3541 tattctttag aatggtgcaa agaataactt cgtataatgt atgctatacg aagttatgaa
     3601 ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     3661 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     3721 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     3781 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     3841 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     3901 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     3961 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     4021 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     4081 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     4141 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     4201 gcctccccgc atcgataccg tcgacctcga cccgggcggc cgcttcgagc agacatgata
     4261 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     4321 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     4381 aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt
     4441 taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatcttcc tagagcatgg
     4501 ctacgtagat aagtagcatg gcgggttaat cattaactac aaggaacccc tagtgatgga
     4561 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     4621 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctggcgtaa
     4681 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     4741 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4801 cgctacacyt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4861 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4921 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggct gtccattaat
     4981 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5041 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5101 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5161 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5221 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5281 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5341 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5401 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5461 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5521 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5581 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5641 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5701 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5761 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5821 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5881 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5941 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6001 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6061 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6121 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6181 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6241 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6301 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6361 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6421 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6481 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6541 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6601 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6661 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6721 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6781 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6841 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6901 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6961 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
     7021 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     7081 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     7141 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     7201 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     7261 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     7321 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     7381 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     7441 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt
     7501 tggg
//
