LOCUS       Exported                6349 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  ReaChR-citrine in AAV2 vector under human synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    AAV-ReaChR-citrine
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6349)
  AUTHORS   Lin JY, Knutsen PM, Muller A, Kleinfeld D, Tsien RY
  TITLE     ReaChR: a red-shifted variant of channelrhodopsin enables deep 
            transcranial optogenetic excitation.
  JOURNAL   Nat Neurosci. 2013 Oct;16(10):1499-508. doi: 10.1038/nn.3502. Epub 
            2013 Sep 1.
  PUBMED    23995068
REFERENCE   2  (bases 1 to 6349)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6349
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     94..111
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    228..249
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        264..294
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    302..318
                     /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     307..329
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     326..342
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     326..342
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   380..509
                     /label=AAV2 ITR
     promoter        591..1038
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             2126..2842
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2171..2192)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2432..2451)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2779..2800
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2858..3446
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2911..2931)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3329..3340)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3449..3465)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3450..3466)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3495..3616
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3506..3525)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3560..3579
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3778..3907
                     /label=AAV2 ITR
     primer_bind     complement(3926..3943)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3926..3942)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3935..3957)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4084..4539
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4171..4190)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4381..4402
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4565..4669
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4670..5530
                     /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(4888..4907)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5701..6289
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6190..6209
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
       61 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      121 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      181 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      241 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      301 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccagattt
      361 aattaaggcc ttaattaggc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc
      421 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
      481 agtggccaac tccatcacta ggggttcctt gtagttaatg attaacccgc catgctactt
      541 atctacgtag ccatgctcta ggaagatctc tgcagagggc cctgcgtatg agtgcaagtg
      601 ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga ccccgaccca
      661 ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga gagggggagg
      721 ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct
      781 tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca
      841 ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc
      901 ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg
      961 cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg
     1021 tcgtgcctga gagcgcagtc gaattcaagc tgctagcaag gatccgacca tggtgagcag
     1081 aagaccctgg ctgctggccc tggccctggc cgtggccctg gccgccggca gcgccggcgc
     1141 cagcaccggc agcgacgcca ccgtgcccgt ggccacccag gacggccccg actacgtgtt
     1201 ccacagagcc cacgagagaa tgctgttcca gaccagctac accctggaga acaacggcag
     1261 cgtgatctgc atccccaaca acggccagtg cttctgcctg gcctggctga agagtaacgg
     1321 caccaacgcc gagaagctgg ccgccaacat cctgcagtgg gtggtgtttg cgctgagcgt
     1381 ggcgtgcctg ggctggtatg cgtatcaggc gtggcgcgcg acctgcggct gggaagaagt
     1441 gtatgtggcg ctgattgaaa tgatgaaaag cattattgaa gcgtttcatg aatttgatag
     1501 cccggcgacc ctgtggctga gcagcggcaa cggcgtggtg tggatgcgct atggcgaatg
     1561 gctgctgacc tgccccgtga ttctgattca tctgagcaac ctgaccggcc tgaaagatga
     1621 ttatagcaaa cgcaccatgg gcctgctggt gagcgacgtg ggctgcattg tgtggggcgc
     1681 gaccagcgcg atgtgcaccg gctggaccaa aattctgttt tttctgatta gcctgagcta
     1741 tggcatgtat acctattttc atgcggccaa agtgtatatt gaagcgtttc ataccgtgcc
     1801 gaaaggcctg tgcagacagc tggtgagagc catggcctgg ctgttcttcg tgagctgggg
     1861 catgttcccc gtgctgttcc tgctgggccc cgagggcttc ggccatatta gcccgtatgg
     1921 cagcgcgatt ggccatagca ttctggatct gattgcgaag aacatgtggg gcgtgctggg
     1981 caactatctg cgcgtgaaaa ttcatgaaca tattctgctg tatggcgata ttcgcaaaaa
     2041 acagaaaatt accattgcgg gccaggaaat ggaagtggaa accctggtgg cggaagaaga
     2101 agataagtac gagagcagcc tcgagatggt gagcaagggc gaggagctgt tcaccggggt
     2161 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     2221 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     2281 caagctgccc gtgccctggc ccaccctcgt gaccaccttc ggctacggcc tgatgtgctt
     2341 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     2401 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     2461 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     2521 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     2581 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     2641 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     2701 ccccgtgctg ctgcccgaca accactacct gagctaccag tccgccctga gcaaagaccc
     2761 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     2821 cggcatggac gagctgtaca agtagaagct tatcgataat caacctctgg attacaaaat
     2881 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     2941 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     3001 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     3061 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     3121 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     3181 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     3241 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     3301 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     3361 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     3421 gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgacccg ggcggccgct
     3481 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     3541 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     3601 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     3661 gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga
     3721 tcttcctaga gcatggctac gtagataagt agcatggcgg gttaatcatt aactacaagg
     3781 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     3841 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     3901 cgcgcagcct taattaacct aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     3961 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     4021 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     4081 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4141 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4201 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4261 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     4321 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4381 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     4441 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4501 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
     4561 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4621 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4681 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4741 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4801 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4861 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4921 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4981 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     5041 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     5101 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5161 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     5221 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     5281 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5341 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5401 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5461 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5521 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     5581 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5641 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5701 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5761 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5821 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5881 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5941 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6001 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6061 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6121 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6181 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6241 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6301 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatg
//
