LOCUS       Exported                6746 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  A somatic channelrhodopsin, which enables single cell, single 
            millisecond resolution optogenetics. Human Synapsin (hSynapsin) 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-hSynapsin- soCoChR-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6746)
  AUTHORS   Shemesh OA, Tanese D, Zampini V, Linghu C, Piatkevich K, Ronzitti E,
            Papagiakoumou E, Boyden ES, Emiliani V
  TITLE     Temporally precise single-cell-resolution optogenetics.
  JOURNAL   Nat Neurosci. 2017 Dec;20(12):1796-1806. doi: 
            10.1038/s41593-017-0018-8. Epub 2017 Nov 13.
  PUBMED    29184208
REFERENCE   2  (bases 1 to 6746)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6746
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR
     polyA_signal    186..234
                     /note="synthetic polyadenylation signal"
     misc_feature    248..339
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     288..307
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        375..822
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             856..1719
                     /codon_start=1
                     /product="high-photocurrent channelrhodopsin from 
                     Chloromonas oogama (Shemesh et al., 2017)"
                     /label=CoChR
                     /translation="MLGNGSAIVPIDQCFCLAWTDSLGSDTEQLVANILQWFAFGFSIL
                     ILMFYAYQTWRATCGWEEVYVCCVELTKVIIEFFHEFDDPSMLYLANGHRVQWLRYAEW
                     LLTCPVILIHLSNLTGLKDDYSKRTMRLLVSDVGTIVWGATSAMSTGYVKVIFFVLGCI
                     YGANTFFHAAKVYIESYHVVPKGRPRTVVRIMAWLFFLSWGMFPVLFVVGPEGFDAISV
                     YGSTIGHTIIDLMSKNCWGLLGHYLRVLIHQHIIIYGDIRKKTKINVAGEEMEVETMVD
                     QEDEETV"
     CDS             1756..2205
                     /codon_start=1
                     /product="first 150 residues of the mouse kainate receptor 
                     subunit 2"
                     /label=KA2
                     /note="mediates targeting to the neuronal soma (Shemesh et 
                     al., 2017)"
                     /translation="MPAELLLLLIVAFANPSCQVLSSLRMAAILDDQTVCGRGERLALA
                     LAREQINGIIEVPAKARVEVDIFELQRDSQYETTDTMCQILPKGVVSVLGPSSSPASAS
                     TVSHICGEKEIPHIKVGPEETPRLQYLRFASVSLYPSNEDVSLAVS"
     CDS             2224..2937
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2266..2287)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2527..2546)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2874..2895
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2953..3541
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3006..3026)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3424..3435)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3574..3591)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3580..3787
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3796..3925
                     /label=AAV2 ITR
     primer_bind     complement(3944..3961)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3944..3960)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3953..3975)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4102..4557
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4189..4208)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4399..4420
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4583..4687
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4688..5548
                     /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(4906..4925)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5719..6307
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6208..6227
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6461..6478
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6595..6616
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6631..6661
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6669..6685
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6674..6696
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6693..6709
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6693..6709
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttctaga aatattaagg tacgggaggt acttggagcg
      181 gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga
      241 tagtactaac atacgctctc catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg
      301 ctgtccccag tgcaagtgca ggtgccagaa catttctcta ttctagactc gagctgcaga
      361 gggccctgcg tatgagtgca agtgggtttt aggaccagga tgaggcgggg tgggggtgcc
      421 tacctgacga ccgaccccga cccactggac aagcacccaa cccccattcc ccaaattgcg
      481 catcccctat cagagagggg gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca
      541 gcttcagcac cgcggacagt gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc
      601 agcactgaag gcgcgctgac gtcactcgcc ggtcccccgc aaactcccct tcccggccac
      661 cttggtcgcg tccgcgccgc cgccggccca gccggaccgc accacgcgag gcgcgagata
      721 ggggggcacg ggcgcgacca tctgcgctgc ggcgccggcg actcagcgct gcctcagtct
      781 gcggtgggca gcggaggagt cgtgtcgtgc ctgagagcgc agtcgaattc aagctgctag
      841 caaggatccg ccaccatgct ggggaacggc agcgccattg tgcctatcga ccagtgcttt
      901 tgcctggctt ggaccgacag cctgggaagc gatacagagc agctggtggc caacatcctc
      961 cagtggttcg ccttcggctt cagcatcctg atcctgatgt tctacgccta ccagacttgg
     1021 agagccactt gcggttggga ggaggtctac gtctgttgcg tcgagctgac caaggtcatc
     1081 atcgagttct tccacgagtt cgacgacccc agcatgctgt acctggctaa cggacaccga
     1141 gtccagtggc tgagatacgc agagtggctg ctgacttgtc ccgtcatcct gatccacctg
     1201 agcaacctga ccggcctgaa ggacgactac agcaagcgga ccatgaggct gctggtgtca
     1261 gacgtgggaa ccatcgtgtg gggagctaca agcgccatga gcacaggcta cgtcaaggtc
     1321 atcttcttcg tgctgggttg catctacggc gccaacacct tcttccacgc cgccaaggtg
     1381 tatatcgaga gctaccacgt ggtgccaaag ggcagaccta gaaccgtcgt gcggatcatg
     1441 gcttggctgt tcttcctgtc ttggggcatg ttccccgtgc tgttcgtcgt gggaccagaa
     1501 ggattcgacg ccatcagcgt gtacggctct accattggcc acaccatcat cgacctcatg
     1561 agcaagaatt gttggggcct gctgggacac tatctgagag tgctgatcca ccagcacatc
     1621 atcatctacg gcgacatccg gaagaagacc aagatcaacg tggccggcga ggagatggaa
     1681 gtggagacca tggtggacca ggaggacgag gagacagtgg gaggttcagg tggaaccggt
     1741 ggaagtggag gtaccatgcc cgccgaactg ctgctgctgc tgattgtcgc ttttgctaac
     1801 ccttcttgcc aggtgctgtc ttctctgagg atggccgcta ttctggacga tcagaccgtg
     1861 tgcggaagag gagagaggct ggcactggca ctggctaggg agcagatcaa tggcatcatt
     1921 gaagtgcctg caaaggcccg ggtcgaggtg gacattttcg aactgcagag agatagccag
     1981 tacgagacca cagacaccat gtgccagatc ctgccaaaag gagtggtctc cgtcctggga
     2041 ccaagctcct ctcctgcttc tgcaagtacc gtgtctcaca tttgtggcga gaaggaaatc
     2101 ccccatatca aggtcgggcc agaggaaaca cccaggctgc agtacctgcg cttcgcctca
     2161 gtgagcctgt atccatcaaa cgaggatgtg tcactggcag tcagcggagc tagcggaggt
     2221 actgtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     2281 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     2341 ggcaagctga ccctgaagtt catttgcacc accggcaagc tgcccgtgcc ctggcccacc
     2401 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     2461 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     2521 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     2581 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     2641 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     2701 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     2761 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     2821 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     2881 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     2941 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     3001 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     3061 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     3121 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     3181 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     3241 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     3301 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     3361 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgccctt ctgctacgtc
     3421 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     3481 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     3541 catcgatacc gtcgactaga gctcgctgat cagcctcgac tgtgccttct agttgccagc
     3601 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     3661 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     3721 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     3781 ctggggagag atctgaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     3841 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     3901 cctcagtgag cgagcgagcg cgcagcctta attaacctaa ttcactggcc gtcgttttac
     3961 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     4021 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     4081 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4141 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4201 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4261 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4321 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4381 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4441 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     4501 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     4561 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     4621 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4681 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4741 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4801 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4861 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4921 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4981 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5041 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5101 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5161 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5221 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     5281 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     5341 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5401 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5461 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     5521 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     5581 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5641 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5701 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5761 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5821 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5881 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5941 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6001 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6061 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6121 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6181 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6241 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6301 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6361 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     6421 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     6481 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     6541 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     6601 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     6661 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     6721 cagatttaat taaggcctta attagg
//
