LOCUS       Exported                6363 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  AAV-mediated expression of ChromeT-GFP under the Syn promoter..
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-Syn-ChromeT-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6363)
  AUTHORS   Cho YK, Park D, Yang A, Chen F, Chuong AS, Klapoetke NC, Boyden ES
  TITLE     Multidimensional screening yields channelrhodopsin variants having 
            improved photocurrent and order-of-magnitude reductions in calcium 
            and proton currents.
  JOURNAL   J Biol Chem. 2019 Jan 4. pii: RA118.006996. doi: 
            10.1074/jbc.RA118.006996.
  PUBMED    30610117
REFERENCE   2  (bases 1 to 6363)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6363
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        341..788
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      801..810
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             807..1733
                     /codon_start=1
                     /product="H134R mutant of an N-terminal fragment of 
                     channelrhodopsin-2 from the alga Chlamydomonas reinhardtii 
                     (Nagel et al., 2005)"
                     /label=ChR2(H134R)
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYSYQTWKSTCGWEEIYVCAIAMVKVILEFFFEFKN
                     PSMLYLATGGRVQWLRYAEWLLTCPVILIHLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     CDS             1755..2468
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1797..1818)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2058..2077)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2405..2426
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2496..3084
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2549..2569)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2967..2978)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3104..3580
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3303..3322)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3626..3766
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3626..3755
                     /label=AAV2 ITR
     rep_origin      3841..4296
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3928..3947)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4138..4159
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4313..4332)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4432..4454
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4492..4510)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4578..4682
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4683..5543
                     /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(4901..4920)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5714..6302
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6203..6222
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttgcaaa gatggataaa gttttaaaca gagaggaatc
      181 tttgcagcta atggaccttc taggtcttga aaggagtggg aattggctcc ggtgcccgtc
      241 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcagca
      301 aatggttaat taatctagac tgcagagggc cctgcgtatg agtgcaagtg ggttttagga
      361 ccaggatgag gcggggtggg ggtgcctacc tgacgaccga ccccgaccca ctggacaagc
      421 acccaacccc cattccccaa attgcgcatc ccctatcaga gagggggagg ggaaacagga
      481 tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct tcgcccccgc
      541 ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca ctcgccggtc
      601 ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc ggcccagccg
      661 gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg cgctgcggcg
      721 ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg tcgtgcctga
      781 gagcgcagtc gagaggatcc gccaccatgg actatggcgg cgctttgtct gccgtcggac
      841 gcgaactttt gttcgttact aatcctgtgg tggtgaacgg gtccgtcctg gtccctgagg
      901 atcaatgtta ctgtgccgga tggattgaat ctcgcggcac gaacggcgct cagaccgcgt
      961 caaatgtcct gcagtggctt gcagcaggat tcagcatttt gctgctgatg ttctattctt
     1021 accaaacctg gaaatctaca tgcggctggg aggagatcta tgtgtgcgcc attgcaatgg
     1081 ttaaggtgat tctcgagttc ttttttgagt ttaagaatcc ctctatgctc taccttgcca
     1141 caggaggtcg ggtgcagtgg ctgcgctatg cagagtggct gctcacttgt cctgtcatcc
     1201 ttatccacct gagcaacctc accggcctga gcaacgacta cagcaggaga accatgggac
     1261 tccttgtctc agacatcggg actatcgtgt ggggggctac cagcgccatg gcaaccggct
     1321 atgttaaagt catcttcttt tgtcttggat tgtgctatgg cgcgaacaca ttttttcacg
     1381 ccgccaaagc atatatcgag ggttatcata ctgtgccaaa gggtcggtgc cgccaggtcg
     1441 tgaccggcat ggcatggctg tttttcgtga gctggggtat gttcccaatt ctcttcattt
     1501 tggggcccga aggttttggc gtcctgagcg tctatggctc caccgtaggt cacacgatta
     1561 ttgatctgat gagtaaaaat tgttgggggt tgttgggaca ctacctgcgc gtcctgatcc
     1621 acgagcacat attgattcac ggagatatcc gcaaaaccac caaactgaac atcggcggaa
     1681 cggagatcga ggtcgagact ctcgtcgaag acgaagccga ggccggagcc gtgccagcgg
     1741 caccggtagt agcagtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1801 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     1861 atgccaccta cggcaagctg accctgaagt tcatttgcac caccggcaag ctgcccgtgc
     1921 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     1981 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     2041 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     2101 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     2161 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     2221 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     2281 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     2341 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     2401 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     2461 tgtacaagta agaattccgt acaaagctta tcgataatca acctctggat tacaaaattt
     2521 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     2581 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     2641 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     2701 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     2761 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     2821 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     2881 tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc tggattctgc
     2941 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     3001 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     3061 tctccctttg ggccgcctcc ccgcatcgat accggctcga gacgggtggc atccctgtga
     3121 cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt
     3181 cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg
     3241 gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg
     3301 ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc
     3361 ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg
     3421 catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc
     3481 caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct
     3541 gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctcgagctga ttttgtaggt
     3601 aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     3661 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     3721 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     3781 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     3841 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3901 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3961 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4021 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     4081 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4141 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     4201 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     4261 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     4321 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     4381 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     4441 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     4501 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     4561 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4621 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4681 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4741 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4801 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4861 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4921 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4981 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5041 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5101 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5161 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5221 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5281 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5341 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5401 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5461 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5521 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5581 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5641 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5701 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5761 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5821 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5881 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5941 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6001 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6061 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6121 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6181 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6241 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6301 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6361 tgt
//
