LOCUS       sequence-489175-        6041 bp    DNA     circular SYN 23-OCT-2025
DEFINITION  ReaChR-citrine in AAV2 vector under human synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6041)
  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 6041)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 23, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1038/nn.3502"; 
            journalName: "Nat Neurosci"; date: "2013-10"; volume: "16"; issue: 
            "10"; pages: "1499-508"
COMMENT     Sequence Label: AAV-ReaChR-citrine
FEATURES             Location/Qualifiers
     source          1..6041
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   39..168
                     /label=AAV2 ITR
     rep_origin      240..695
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(327..346)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     537..558
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        721..825
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(826..894,895..1686)
                     /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(1044..1063)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1857..2445
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2346..2365
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2682..2811
                     /label=AAV2 ITR
     polyA_signal    2973..3094
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3010..3029)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3064..3083
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3123..3139
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3124..3140
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3143..3731
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3249..3260
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3658..3678
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3747..4457,4458..4460,4461..4463))
                     /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(3789..3810)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4138..4157
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4397..4418
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(5551..5998)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
ORIGIN
        1 cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt
       61 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
      121 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct ggcgtaatag
      181 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
      241 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
      301 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
      361 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
      421 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
      481 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
      541 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
      601 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
      661 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
      721 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
      781 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
      841 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
      901 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
      961 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     1021 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     1081 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     1141 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     1201 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     1261 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     1321 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     1381 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     1441 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     1501 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     1561 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     1621 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     1681 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     1741 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     1801 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1861 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1921 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1981 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     2041 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     2101 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     2161 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     2221 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     2281 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     2341 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2401 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2461 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2521 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     2581 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     2641 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctgcgcgct cgctcgctca
     2701 ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga
     2761 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc ttgtagttaa
     2821 tgattaaccc gccatgctac ttatctacgt agccatgctc taggaagatc cttatcgatt
     2881 ttaccacatt tgtagaggtt ttacttgctt taaaaaacct cccacatctc cccctgaacc
     2941 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     3001 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3061 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgctcgaag cggccgcccg
     3121 ggtcgaggtc gacggtatcg atgcggggag gcggcccaaa gggagatccg actcgtctga
     3181 gggcgaaggc gaagacgcgg aagaggccgc agagccggca gcaggccgcg ggaaggaagg
     3241 tccgctggat tgagggccga agggacgtag cagaaggacg tcccgcgcag aatccaggtg
     3301 gcaacacagg cgagcagcca aggaaaggac gatgatttcc ccgacaacac cacggaattg
     3361 tcagtgccca acagccgagc ccctgtccag cagcgggcaa ggcaggcggc gatgagttcc
     3421 gccgtggcaa tagggagggg gaaagcgaaa gtcccggaaa ggagctgaca ggtggtggca
     3481 atgccccaac cagtgggggt tgcgtcagca aacacagtgc acaccacgcc acgttgcctg
     3541 acaacgggcc acaactcctc ataaagagac agcaaccagg atttatacaa ggaggagaaa
     3601 atgaaagcca tacgggaagc aatagcatga tacaaaggca ttaaagcagc gtatccacat
     3661 agcgtaaaag gagcaacata gttaagaata ccagtcaatc tttcacaaat tttgtaatcc
     3721 agaggttgat tatcgataag cttctacttg tacagctcgt ccatgccgag agtgatcccg
     3781 gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc
     3841 agggcggact ggtagctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg
     3901 ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg
     3961 atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg
     4021 ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
     4081 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
     4141 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg
     4201 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcgggcgaa gcacatcagg
     4261 ccgtagccga aggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
     4321 atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg
     4381 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
     4441 agctcctcgc ccttgctcac catctcgagg ctgctctcgt acttatcttc ttcttccgcc
     4501 accagggttt ccacttccat ttcctggccc gcaatggtaa ttttctgttt tttgcgaata
     4561 tcgccataca gcagaatatg ttcatgaatt ttcacgcgca gatagttgcc cagcacgccc
     4621 cacatgttct tcgcaatcag atccagaatg ctatggccaa tcgcgctgcc atacgggcta
     4681 atatggccga agccctcggg gcccagcagg aacagcacgg ggaacatgcc ccagctcacg
     4741 aagaacagcc aggccatggc tctcaccagc tgtctgcaca ggcctttcgg cacggtatga
     4801 aacgcttcaa tatacacttt ggccgcatga aaataggtat acatgccata gctcaggcta
     4861 atcagaaaaa acagaatttt ggtccagccg gtgcacatcg cgctggtcgc gccccacaca
     4921 atgcagccca cgtcgctcac cagcaggccc atggtgcgtt tgctataatc atctttcagg
     4981 ccggtcaggt tgctcagatg aatcagaatc acggggcagg tcagcagcca ttcgccatag
     5041 cgcatccaca ccacgccgtt gccgctgctc agccacaggg tcgccgggct atcaaattca
     5101 tgaaacgctt caataatgct tttcatcatt tcaatcagcg ccacatacac ttcttcccag
     5161 ccgcaggtcg cgcgccacgc ctgatacgca taccagccca ggcacgccac gctcagcgca
     5221 aacaccaccc actgcaggat gttggcggcc agcttctcgg cgttggtgcc gttactcttc
     5281 agccaggcca ggcagaagca ctggccgttg ttggggatgc agatcacgct gccgttgttc
     5341 tccagggtgt agctggtctg gaacagcatt ctctcgtggg ctctgtggaa cacgtagtcg
     5401 gggccgtcct gggtggccac gggcacggtg gcgtcgctgc cggtgctggc gccggcgctg
     5461 ccggcggcca gggccacggc cagggccagg gccagcagcc agggtcttct gctcaccatg
     5521 gtcggatcct tgctagcagc ttgaattcga ctgcgctctc aggcacgaca cgactcctcc
     5581 gctgcccacc gcagactgag gcagcgctga gtcgccggcg ccgcagcgca gatggtcgcg
     5641 cccgtgcccc cctatctcgc gcctcgcgtg gtgcggtccg gctgggccgg cggcggcgcg
     5701 gacgcgacca aggtggccgg gaaggggagt ttgcggggga ccggcgagtg acgtcagcgc
     5761 gccttcagtg ctgaggcggc ggtggcgcgc gccgccaggc gggggcgaag gcactgtccg
     5821 cggtgctgaa gctggcagtg cgcacgcgcc tcgccgcatc ctgtttcccc tccccctctc
     5881 tgatagggga tgcgcaattt ggggaatggg ggttgggtgc ttgtccagtg ggtcggggtc
     5941 ggtcgtcagg taggcacccc caccccgcct catcctggtc ctaaaaccca cttgcactca
     6001 tacgcagggc cctctgcaga gatcttccta gagcatggct a
//
