LOCUS       Exported                7249 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  vCre-Dependent ChR2-EYFP.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-Ef1a-vCreDIO hChR2(H134R)-EYFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7249)
  AUTHORS   Fenno LE, Mattis J, Ramakrishnan C, Hyun M, Lee SY, He M, Tucciarone
            J, Selimbeyoglu A, Berndt A, Grosenick L, Zalocusky KA, Bernstein H,
            Swanson H, Perry C, Diester I, Boyce FM, Bass CE, Neve R, Huang ZJ, 
            Deisseroth K
  TITLE     Targeting cells with single vectors using multiple-feature Boolean 
            logic.
  JOURNAL   Nat Methods. 2014 Jul;11(7):763-72. doi: 10.1038/nmeth.2996. Epub 
            2014 Jun 8.
  PUBMED    24908100
REFERENCE   2  (bases 1 to 7249)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7249
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        242..1419
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          473..1410
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1367..1387
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             complement(1562..2281)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1607..1628)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1956..1975
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2215..2236
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2278..2287
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             complement(2297..3223)
                     /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
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIRLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     regulatory      3220..3229
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    3376..3964
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3429..3449)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3847..3858)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3996..4472
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4195..4214)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4512..4652
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4512..4641
                     /label=AAV2 ITR
     rep_origin      4727..5182
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4814..4833)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5024..5045
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5199..5218)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5318..5340
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5378..5396)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5464..5568
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5569..6429
                     /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(5787..5806)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6600..7188
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7089..7108
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc catcaatttc
     1441 tgagaactgt cattctcgga aattgagtag ctggagcaat ttgaaaagat tgttgtatta
     1501 cctctagctg cgcttatcaa tttctgagaa gtgtctttct cggaaattga ggcgcgccct
     1561 attacttgta cagctcgtcc atgccgagag tgatcccggc ggcggtcacg aactccagca
     1621 ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag ggcggactgg tagctcaggt
     1681 agtggttgtc gggcagcagc acggggccgt cgccgatggg ggtgttctgc tggtagtggt
     1741 cggcgagctg cacgctgccg tcctcgatgt tgtggcggat cttgaagttc accttgatgc
     1801 cgttcttctg cttgtcggcc atgatataga cgttgtggct gttgtagttg tactccagct
     1861 tgtgccccag gatgttgccg tcctccttga agtcgatgcc cttcagctcg atgcggttca
     1921 ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga
     1981 agaagatggt gcgctcctgg acgtagcctt cgggcatggc ggacttgaag aagtcgtgct
     2041 gcttcatgtg gtcggggtag cgggcgaagc actgcaggcc gtagccgaag gtggtcacga
     2101 gggtgggcca gggcacgggc agcttgccgg tggtgcagat gaacttcagg gtcagcttgc
     2161 cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc
     2221 cgtccagctc gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca
     2281 tggtggcggc cgctggcacg gctccggcct cggcttcgtc ttcgacgaga gtctcgacct
     2341 cgatctccgt tccgccgatg ttcagtttgg tggttttgcg gatatctccg tgaatcaata
     2401 tgtgctcgtg gatcaggacg cgcaggtagt gtcccaacaa cccccaacaa tttttactca
     2461 tcagatcaat aatcgtgtga cctacggtgg agccatagac gctcaggacg ccaaaacctt
     2521 cgggccccaa aatgaagaga attgggaaca taccccagct cacgaaaaac agccatgcca
     2581 tgccggtcac gacctggcgg caccgaccct ttggcacagt atgataaccc tcgatatatg
     2641 ctttggcggc gtgaaaaaat gtgttcgcgc catagcacaa tccaagacaa aagaagatga
     2701 ctttaacata gccggttgcc atggcgctgg tagcccccca cacgatagtc ccgatgtctg
     2761 agacaaggag tcccatggtt ctcctgctgt agtcgttgct caggccggtg aggttgctca
     2821 ggcggataag gatgacagga caagtgagca gccactctgc atagcgcagc cactgcaccc
     2881 ggtgtcctgt ggcaaggtag agcatagagg gattcttaaa ctcaaaaaag aactcgagaa
     2941 tcaccttaac catttcaatg gcgcacacat agatctcctc ccagccgcat gtagatttcc
     3001 aggtttggta ggcatagaac atcagcagca aaatgctgaa tcctgctgca agccactgca
     3061 ggacatttga cgcggtctga gcgccgttcg tgccgcgaga ttcaatccat ccggcacagt
     3121 aacattgatc ctcagggacc aggacggacc cgttcaccac cacaggatta gtaacgaaca
     3181 aaagttcgcg tccgacggca gacaaagcgc cgccatagtc catggtggct agctcaattt
     3241 ccgagaaaga cacttctcag aaattgataa gcgcagctag aggtaataca acaatctttt
     3301 caaattgctc cagctactca atttccgaga atgacagttc tcagaaattg agaattcgat
     3361 atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     3421 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     3481 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     3541 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     3601 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     3661 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     3721 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     3781 tttccttggc tgctcgccta tgttgccacc tggattctgc gcgggacgtc cttctgctac
     3841 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     3901 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     3961 ccgcatcgat accgagcgct gctcgagaga tctacgggtg gcatccctgt gacccctccc
     4021 cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa
     4081 aattaagttg catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg
     4141 gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt
     4201 gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg
     4261 ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca
     4321 ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg
     4381 tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac
     4441 aggcgtgaac cactgctccc ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac
     4501 cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     4561 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     4621 agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac
     4681 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
     4741 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4801 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4861 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4921 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     4981 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     5041 actggaacaa cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg
     5101 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     5161 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
     5221 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     5281 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     5341 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     5401 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     5461 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     5521 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     5581 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5641 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     5701 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     5761 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     5821 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     5881 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     5941 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     6001 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     6061 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     6121 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     6181 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     6241 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     6301 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     6361 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     6421 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     6481 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     6541 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     6601 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6661 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6721 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     6781 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6841 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6901 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6961 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     7021 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     7081 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     7141 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     7201 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
