LOCUS       Exported                7058 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  A somatic channelrhodopsin, which enables single cell, single 
            millisecond resolution optogenetics. EF1alpha promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-EF1alpha-soCoChR-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7058)
  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 7058)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7058
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   35..164
                     /label=AAV2 ITR
     promoter        237..1414
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          468..1405
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1362..1382
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1427..2290
                     /codon_start=1
                     /product="high-photocurrent channelrhodopsin from 
                     Chloromonas oogama (Shemesh et al., 2017)"
                     /label=CoChR
                     /translation="MLGNGSAIVPIDQCFCLAWTDSLGSDTEQLVANILQWFAFGFSIL
                     ILMFYAYQTWRATCGWEEVYVCCVELTKVIIEFFHEFDDPSMLYLANGHRVQWLRYAEW
                     LLTCPVILIHLSNLTGLKDDYSKRTMRLLVSDVGTIVWGATSAMSTGYVKVIFFVLGCI
                     YGANTFFHAAKVYIESYHVVPKGRPRTVVRIMAWLFFLSWGMFPVLFVVGPEGFDAISV
                     YGSTIGHTIIDLMSKNCWGLLGHYLRVLIHQHIIIYGDIRKKTKINVAGEEMEVETMVD
                     QEDEETV"
     CDS             2327..2776
                     /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             2795..3508
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2837..2858)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3098..3117)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3445..3466
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3536..4124
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3589..3609)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4007..4018)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4167..4288
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4178..4197)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4232..4251
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4450..4579
                     /label=AAV2 ITR
     primer_bind     complement(4645..4662)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4816..5404)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4896..4915)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5575..6435)
                     /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     6198..6217
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6436..6540)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6566..7021)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6703..6724)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6915..6934
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcgatcggtg cgggcctctt cgctattacg ccagctgcgc gctcgctcgc tcactgaggc
       61 cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg
      121 agcgcgcaga gagggagtgg ccaactccat cactaggggt tccttgtagt taatgattaa
      181 cccgccatgc tacttatcta cgtagccatg ctctaggaag atcgtaggtt aattaaggct
      241 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
      301 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
      361 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
      421 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt
      481 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt
      541 ccactggctg cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag
      601 ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct tgagttgagg cctggcctgg
      661 gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga
      721 taagtctcta gccatttaaa atttttgatg acctgctgcg acgctttttt tctggcaaga
      781 tagtcttgta aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg
      841 cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg
      901 gccaccgaga atcggacggg ggtagtctca agctggccgg cctgctctgg tgcctggcct
      961 cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg caccagttgc
     1021 gtgagcggaa agatggccgc ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg
     1081 gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc
     1141 agccgtcgct tcatgtgact ccacggagta ccgggcgccg tccaggcacc tcgattagtt
     1201 ctcgagcttt tggagtacgt cgtctttagg ttggggggag gggttttatg cgatggagtt
     1261 tccccacact gagtgggtgg agactgaagt taggccagct tggcacttga tgtaattctc
     1321 cttggaattt gccctttttg agtttggatc ttggttcatt ctcaagcctc agacagtggt
     1381 tcaaagtttt tttcttccat ttcaggtgtc gtgaggatcc gccaccatgc tggggaacgg
     1441 cagcgccatt gtgcctatcg accagtgctt ttgcctggct tggaccgaca gcctgggaag
     1501 cgatacagag cagctggtgg ccaacatcct ccagtggttc gccttcggct tcagcatcct
     1561 gatcctgatg ttctacgcct accagacttg gagagccact tgcggttggg aggaggtcta
     1621 cgtctgttgc gtcgagctga ccaaggtcat catcgagttc ttccacgagt tcgacgaccc
     1681 cagcatgctg tacctggcta acggacaccg agtccagtgg ctgagatacg cagagtggct
     1741 gctgacttgt cccgtcatcc tgatccacct gagcaacctg accggcctga aggacgacta
     1801 cagcaagcgg accatgaggc tgctggtgtc agacgtggga accatcgtgt ggggagctac
     1861 aagcgccatg agcacaggct acgtcaaggt catcttcttc gtgctgggtt gcatctacgg
     1921 cgccaacacc ttcttccacg ccgccaaggt gtatatcgag agctaccacg tggtgccaaa
     1981 gggcagacct agaaccgtcg tgcggatcat ggcttggctg ttcttcctgt cttggggcat
     2041 gttccccgtg ctgttcgtcg tgggaccaga aggattcgac gccatcagcg tgtacggctc
     2101 taccattggc cacaccatca tcgacctcat gagcaagaat tgttggggcc tgctgggaca
     2161 ctatctgaga gtgctgatcc accagcacat catcatctac ggcgacatcc ggaagaagac
     2221 caagatcaac gtggccggcg aggagatgga agtggagacc atggtggacc aggaggacga
     2281 ggagacagtg ggaggttcag gtggaaccgg tggaagtgga ggtaccatgc ccgccgaact
     2341 gctgctgctg ctgattgtcg cttttgctaa cccttcttgc caggtgctgt cttctctgag
     2401 gatggccgct attctggacg atcagaccgt gtgcggaaga ggagagaggc tggcactggc
     2461 actggctagg gagcagatca atggcatcat tgaagtgcct gcaaaggccc gggtcgaggt
     2521 ggacattttc gaactgcaga gagatagcca gtacgagacc acagacacca tgtgccagat
     2581 cctgccaaaa ggagtggtct ccgtcctggg accaagctcc tctcctgctt ctgcaagtac
     2641 cgtgtctcac atttgtggcg agaaggaaat cccccatatc aaggtcgggc cagaggaaac
     2701 acccaggctg cagtacctgc gcttcgcctc agtgagcctg tatccatcaa acgaggatgt
     2761 gtcactggca gtcagcggag ctagcggagg tactgtgagc aagggcgagg agctgttcac
     2821 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     2881 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatttgcac
     2941 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     3001 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     3061 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     3121 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     3181 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     3241 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     3301 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     3361 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     3421 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     3481 cactctcggc atggacgagc tgtacaagta agaattcgat atcaagctta tcgataatca
     3541 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     3601 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     3661 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     3721 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     3781 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     3841 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     3901 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     3961 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     4021 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     4081 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc
     4141 cgggcggccg cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac
     4201 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     4261 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     4321 ggttcagggg gagatgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa
     4381 aatcgataag gatcttccta gagcatggct acgtagataa gtagcatggc gggttaatca
     4441 ttaactacaa ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     4501 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     4561 tgagcgagcg agcgcgcagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4621 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4681 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4741 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4801 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4861 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4921 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4981 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5041 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5101 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5161 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5221 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5281 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5341 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5401 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5461 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5521 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5581 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5641 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5701 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5761 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5821 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5881 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5941 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6001 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6061 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6121 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6181 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6241 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6301 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6361 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6421 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6481 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6541 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
     6601 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     6661 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     6721 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     6781 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     6841 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     6901 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     6961 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     7021 tcccattcgc cattcaggct gcgcaactgt tgggaagg
//
