LOCUS       Exported                6689 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  A somatic channelrhodopsin, which enables single cell, single 
            millisecond resolution optogenetics. hSynapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-hSyn-soCoChR-mCardinal
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6689)
  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 6689)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6689
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   58..187
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     polyA_signal    243..291
                     /note="synthetic polyadenylation signal"
     misc_feature    305..396
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     345..364
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        432..879
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             913..1776
                     /codon_start=1
                     /product="high-photocurrent channelrhodopsin from 
                     Chloromonas oogama (Shemesh et al., 2017)"
                     /label=CoChR
                     /translation="MLGNGSAIVPIDQCFCLAWTDSLGSDTEQLVANILQWFAFGFSIL
                     ILMFYAYQTWRATCGWEEVYVCCVELTKVIIEFFHEFDDPSMLYLANGHRVQWLRYAEW
                     LLTCPVILIHLSNLTGLKDDYSKRTMRLLVSDVGTIVWGATSAMSTGYVKVIFFVLGCI
                     YGANTFFHAAKVYIESYHVVPKGRPRTVVRIMAWLFFLSWGMFPVLFVVGPEGFDAISV
                     YGSTIGHTIIDLMSKNCWGLLGHYLRVLIHQHIIIYGDIRKKTKINVAGEEMEVETMVD
                     QEDEETV"
     CDS             1813..2262
                     /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             2281..3012
                     /codon_start=1
                     /product="far-red fluorescent protein derived from 
                     mNeptune, with further red-shifted spectra"
                     /label=mCardinal
                     /translation="VSKGEELIKENMHMKLYMEGTVNNHHFKCTTEGEGKPYEGTQTQR
                     IKVVEGGPLPFAFDILATCFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGV
                     LTVTQDTSLQDGCLIYNVKLRGVNFPSNGPVMQKKTLGWEATTETLYPADGGLEGRCDM
                     ALKLVGGGHLHCNLKTTYRSKKPAKNLKMPGVYFVDRRLERIKEADNETYVEQHEVAVA
                     RYCDLPSKLGHKLNGMDELYK"
     misc_feature    3025..3613
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3078..3098)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3496..3507)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3646..3663)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3652..3859
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3868..3997
                     /label=AAV2 ITR
     primer_bind     complement(4016..4033)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4016..4032)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4025..4047)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4174..4629
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4261..4280)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4471..4492
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4696..4715)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4815..4837
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4875..4893)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4961..5065
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5066..5926
                     /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(5284..5303)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6097..6685
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6586..6605
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtcct
       61 gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt
      121 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg
      181 ggttcctgcg gccgcacgcg ttctagaaat attaaggtac gggaggtact tggagcggcc
      241 gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag
      301 tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca aaataggctg
      361 tccccagtgc aagtgcaggt gccagaacat ttctctattc tagactcgag ctgcagaggg
      421 ccctgcgtat gagtgcaagt gggttttagg accaggatga ggcggggtgg gggtgcctac
      481 ctgacgaccg accccgaccc actggacaag cacccaaccc ccattcccca aattgcgcat
      541 cccctatcag agagggggag gggaaacagg atgcggcgag gcgcgtgcgc actgccagct
      601 tcagcaccgc ggacagtgcc ttcgcccccg cctggcggcg cgcgccaccg ccgcctcagc
      661 actgaaggcg cgctgacgtc actcgccggt cccccgcaaa ctccccttcc cggccacctt
      721 ggtcgcgtcc gcgccgccgc cggcccagcc ggaccgcacc acgcgaggcg cgagataggg
      781 gggcacgggc gcgaccatct gcgctgcggc gccggcgact cagcgctgcc tcagtctgcg
      841 gtgggcagcg gaggagtcgt gtcgtgcctg agagcgcagt cgaattcaag ctgctagcaa
      901 ggatccgcca ccatgctggg gaacggcagc gccattgtgc ctatcgacca gtgcttttgc
      961 ctggcttgga ccgacagcct gggaagcgat acagagcagc tggtggccaa catcctccag
     1021 tggttcgcct tcggcttcag catcctgatc ctgatgttct acgcctacca gacttggaga
     1081 gccacttgcg gttgggagga ggtctacgtc tgttgcgtcg agctgaccaa ggtcatcatc
     1141 gagttcttcc acgagttcga cgaccccagc atgctgtacc tggctaacgg acaccgagtc
     1201 cagtggctga gatacgcaga gtggctgctg acttgtcccg tcatcctgat ccacctgagc
     1261 aacctgaccg gcctgaagga cgactacagc aagcggacca tgaggctgct ggtgtcagac
     1321 gtgggaacca tcgtgtgggg agctacaagc gccatgagca caggctacgt caaggtcatc
     1381 ttcttcgtgc tgggttgcat ctacggcgcc aacaccttct tccacgccgc caaggtgtat
     1441 atcgagagct accacgtggt gccaaagggc agacctagaa ccgtcgtgcg gatcatggct
     1501 tggctgttct tcctgtcttg gggcatgttc cccgtgctgt tcgtcgtggg accagaagga
     1561 ttcgacgcca tcagcgtgta cggctctacc attggccaca ccatcatcga cctcatgagc
     1621 aagaattgtt ggggcctgct gggacactat ctgagagtgc tgatccacca gcacatcatc
     1681 atctacggcg acatccggaa gaagaccaag atcaacgtgg ccggcgagga gatggaagtg
     1741 gagaccatgg tggaccagga ggacgaggag acagtgggag gttcaggtgg aaccggtgga
     1801 agtggaggta ccatgcccgc cgaactgctg ctgctgctga ttgtcgcttt tgctaaccct
     1861 tcttgccagg tgctgtcttc tctgaggatg gccgctattc tggacgatca gaccgtgtgc
     1921 ggaagaggag agaggctggc actggcactg gctagggagc agatcaatgg catcattgaa
     1981 gtgcctgcaa aggcccgggt cgaggtggac attttcgaac tgcagagaga tagccagtac
     2041 gagaccacag acaccatgtg ccagatcctg ccaaaaggag tggtctccgt cctgggacca
     2101 agctcctctc ctgcttctgc aagtaccgtg tctcacattt gtggcgagaa ggaaatcccc
     2161 catatcaagg tcgggccaga ggaaacaccc aggctgcagt acctgcgctt cgcctcagtg
     2221 agcctgtatc catcaaacga ggatgtgtca ctggcagtca gcggagctag cggaggtact
     2281 gtgagcaagg gcgaggagct gatcaaggag aacatgcaca tgaagctgta catggaaggc
     2341 accgtgaaca accaccactt caagtgcacc accgaagggg agggcaagcc ctacgagggc
     2401 acccagaccc agaggattaa ggtggtggag ggaggccccc tgccgttcgc attcgacatc
     2461 ctggccacct gctttatgta cgggagcaag accttcatca accacaccca gggcatcccc
     2521 gatttcttta agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa
     2581 gacgggggcg tgcttaccgt tacccaggac accagcctcc aggacggctg cttgatctac
     2641 aacgtcaagc tcagaggggt gaacttccca tccaacggcc ctgtgatgca gaagaaaaca
     2701 ctcggctggg aggccaccac cgagaccctg taccccgctg acggcggcct ggaaggcaga
     2761 tgcgacatgg ccctgaagct cgtgggcggg ggccacctgc actgcaacct gaagaccaca
     2821 tacagatcca agaaacccgc taagaacctc aagatgcccg gcgtctactt tgtggaccgc
     2881 agactggaaa gaatcaagga ggccgacaat gagacctacg tcgagcagca cgaggtggct
     2941 gtggccagat actgcgacct ccctagcaaa ctggggcaca aacttaatgg catggacgag
     3001 ctgtacaagt aaaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     3061 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     3121 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     3181 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     3241 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     3301 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     3361 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     3421 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgccc
     3481 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     3541 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     3601 gccgcctccc cgcatcgata ccgtcgacta gagctcgctg atcagcctcg actgtgcctt
     3661 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     3721 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     3781 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     3841 atagcaggca tgctggggag agatctgagg aacccctagt gatggagttg gccactccct
     3901 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     3961 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagcct taattaacct aattcactgg
     4021 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     4081 cagcacatcc ccctttcgcc agcctggcgt aatagcgaag aggcccgcac cgatcgccct
     4141 tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg
     4201 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     4261 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4321 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4381 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     4441 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     4501 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     4561 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     4621 ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     4681 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     4741 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     4801 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     4861 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     4921 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4981 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5041 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5101 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5161 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5221 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5281 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5341 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5401 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5461 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5521 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5581 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5641 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5701 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5761 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5821 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5881 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     5941 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6001 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6061 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6121 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6181 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6241 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6301 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6361 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6421 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6481 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6541 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6601 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6661 gtcagggggg cggagcctat ggaaaaacg
//
