LOCUS       Exported                6630 bp ds-DNA     circular SYN 16-JAN-2024
DEFINITION  flexed-ReaChR-citrine in AAV2 vector under human synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    AAV-flex-ReaChR-citrine
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6630)
  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 6630)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 16, 2024 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6630
                     /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..1037
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    1083..1116
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    1167..1200
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(1215..1931)
                     /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(1257..1278)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1606..1625
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1865..1886
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(2997..3030)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    complement(3081..3114)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3139..3727
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3192..3212)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3610..3621)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3730..3746)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3731..3747)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3776..3897
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3787..3806)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3841..3860
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4059..4188
                     /label=AAV2 ITR
     primer_bind     complement(4207..4224)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4207..4223)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4216..4238)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4365..4820
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4452..4471)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4662..4683
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4846..4950
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4951..5811
                     /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(5169..5188)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5982..6570
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6471..6490
                     /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 ccccacccac
      661 tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag agggggaggg
      721 gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg acagtgcctt
      781 cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg ctgacgtcac
      841 tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc gccgccgccg
      901 gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc gaccatctgc
      961 gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga ggagtcgtgt
     1021 cgtgcctgag agcgcagtcg aattcaagct gatagcaagg atcctctaga gtcgactccg
     1081 gaataacttc gtataggata ctttatacga agttatgcag aatggtagct ggattgtagc
     1141 tgctattagc aatatgaaac ctcttaataa cttcgtatag catacattat acgaagttat
     1201 ggcgcgccct actacttgta cagctcgtcc atgccgagag tgatcccggc ggcggtcacg
     1261 aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag ggcggactgg
     1321 tagctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg ggtgttctgc
     1381 tggtagtggt cggcgagctg cacgctgccg tcctcgatgt tgtggcggat cttgaagttc
     1441 accttgatgc cgttcttctg cttgtcggcc atgatataga cgttgtggct gttgtagttg
     1501 tactccagct tgtgccccag gatgttgccg tcctccttga agtcgatgcc cttcagctcg
     1561 atgcggttca ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt gtagttgccg
     1621 tcgtccttga agaagatggt gcgctcctgg acgtagcctt cgggcatggc ggacttgaag
     1681 aagtcgtgct gcttcatgtg gtcggggtag cgggcgaagc acatcaggcc gtagccgaag
     1741 gtggtcacga gggtgggcca gggcacgggc agcttgccgg tggtgcagat gaacttcagg
     1801 gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa cttgtggccg
     1861 tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc
     1921 ttgctcacca tctcgaggct gctctcgtac ttatcttctt cttccgccac cagggtttcc
     1981 acttccattt cctggcccgc aatggtaatt ttctgttttt tgcgaatatc gccatacagc
     2041 agaatatgtt catgaatttt cacgcgcaga tagttgccca gcacgcccca catgttcttc
     2101 gcaatcagat ccagaatgct atggccaatc gcgctgccat acgggctaat atggccgaag
     2161 ccctcggggc ccagcaggaa cagcacgggg aacatgcccc agctcacgaa gaacagccag
     2221 gccatggctc tcaccagctg tctgcacagg cctttcggca cggtatgaaa cgcttcaata
     2281 tacactttgg ccgcatgaaa ataggtatac atgccatagc tcaggctaat cagaaaaaac
     2341 agaattttgg tccagccggt gcacatcgcg ctggtcgcgc cccacacaat gcagcccacg
     2401 tcgctcacca gcaggcccat ggtgcgtttg ctataatcat ctttcaggcc ggtcaggttg
     2461 ctcagatgaa tcagaatcac ggggcaggtc agcagccatt cgccatagcg catccacacc
     2521 acgccgttgc cgctgctcag ccacagggtc gccgggctat caaattcatg aaacgcttca
     2581 ataatgcttt tcatcatttc aatcagcgcc acatacactt cttcccagcc gcaggtcgcg
     2641 cgccacgcct gatacgcata ccagcccagg cacgccacgc tcagcgcaaa caccacccac
     2701 tgcaggatgt tggcggccag cttctcggcg ttggtgccgt tactcttcag ccaggccagg
     2761 cagaagcact ggccgttgtt ggggatgcag atcacgctgc cgttgttctc cagggtgtag
     2821 ctggtctgga acagcattct ctcgtgggct ctgtggaaca cgtagtcggg gccgtcctgg
     2881 gtggccacgg gcacggtggc gtcgctgccg gtgctggcgc cggcgctgcc ggcggccagg
     2941 gccacggcca gggccagggc cagcagccag ggtcttctgc tcaccatggt gctagcataa
     3001 cttcgtataa agtatcctat acgaagttat ttgccttaac ccagaaatta tcactgttat
     3061 tctttagaat ggtgcaaaga ataacttcgt ataatgtatg ctatacgaag ttatgaattc
     3121 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     3181 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     3241 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     3301 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     3361 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     3421 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3481 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     3541 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     3601 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     3661 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     3721 tccccgcatc gataccgtcg acctcgaccc gggcggccgc ttcgagcaga catgataaga
     3781 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     3841 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     3901 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa
     3961 agcaagtaaa acctctacaa atgtggtaaa atcgataagg atcttcctag agcatggcta
     4021 cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt
     4081 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     4141 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagcc ttaattaacc
     4201 taattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     4261 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     4321 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
     4381 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4441 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4501 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4561 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4621 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4681 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     4741 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     4801 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4861 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4921 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4981 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5041 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5101 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5161 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5221 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5281 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5341 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5401 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5461 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5521 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5581 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5641 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5701 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5761 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5821 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5881 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5941 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6001 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6061 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6121 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6181 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6241 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6301 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6361 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6421 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6481 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6541 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6601 ctggcctttt gctggccttt tgctcacatg
//
