LOCUS       sequence-355552-        7422 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Opto-cytTrkB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7422)
  AUTHORS   Hong J , Heo WD
  TITLE     Optogenetic Modulation of TrkB Signaling in the Mouse Brain.
  JOURNAL   J Mol Biol. 2020 Feb 14;432(4):815-827. doi: 
            10.1016/j.jmb.2020.01.010. Epub 2020 Jan 18.
  PUBMED    31962123
REFERENCE   2  (bases 1 to 7422)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.jmb.2020.01.010"; journalName: "J Mol Biol"; date: 
            "2020-02-14- 14"; volume: "432"; issue: "4"; pages: "815-827"
COMMENT     Sequence Label: pAAV-Camk2a(0.4)-DIO-Opto-cytTrkB(E281A)-HA
FEATURES             Location/Qualifiers
     source          1..7422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   2..131
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     protein_bind    complement(529..562)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(613..646)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(658..684)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      3393..3402
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3408..3441
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    3492..3525
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3550..4138
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3603..3623)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4021..4032)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4170..4646
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4369..4388)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4686..4826
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4686..4815
                     /label=AAV2 ITR
     rep_origin      4901..5356
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4988..5007)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5198..5219
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5373..5392)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5492..5514
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5552..5570)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5638..5742
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5743..5811,5812..6603)
                     /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(5961..5980)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6774..7362
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7263..7282
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt
       61 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
      121 taggggttcc tgcggccgca cgcgtaactt gtggactaag tttgttcaca tccccttctc
      181 caaccccctc agtacatcac cctgggggaa cagggtccac ttgctcctgg gcccacacag
      241 tcctgcagta ttgtgtatat aaggccaggg caaagaggag caggttttaa agtgaaaggc
      301 aggcaggtgt tggggaggca gttaccgggg caacgggaac agggcgtttc ggaggtggtt
      361 gccatgggga cctggatgct gacgaaggct cgcgaggctg tgagcagcca cagtgccctg
      421 ctcagaagcc ccaagctcgt cagtcaagcc ggttctccgt ttgcactcag gagcacgggc
      481 aggcgagtgg cccctagttc tgggggcagc tctagagtcg actccggaat aacttcgtat
      541 aggatacttt atacgaagtt atgcagaatg gtagctggat tgtagctgct attagcaata
      601 tgaaacctct taataacttc gtatagcata cattatacga agttatggcg cgccttaagc
      661 ataatctgga acatcatatg gatataccgg tgggctgccg ctgccggcag caccgatcat
      721 aatctgcgcc tcccgagtgc gagagatcgc tttagccagc agctcgcgtg ctgtatctat
      781 atcaactata ggcttagcgt aattggtgcc cagctccacc ccgcttgcct tgagcactgt
      841 cagtggtgcg tcccaaggat ggtgaatcca ctccgtagga agtcgggcca gctcggggag
      901 ccactgtcga atatactctc cttcgggatc atacttagct ccctgcaagg ccggattgtc
      961 caacctgtcc aactcatggc cgtcaggaat tgacccgcta atatattgcc atcccagaat
     1021 gtcacactcc aagtcggcgt cgagaagggt atcccaaaag tacttcattc cccatttcca
     1081 aggcaggagc agaaacttga cagcaaatga ggacacgatt acccggatgc ggttgtgcat
     1141 ccagcctgta gcccagagct ctctcatgcc agcatccacg agtgggtacc ctgtccttcc
     1201 ctgtctccat gccttgaatt tgtcgacatc ggcgtcccag gggaagaacc tgaggtggga
     1261 caacagactc tgctcgtgtg tgaatggaaa gttgaaacat atataccggg agtattcccg
     1321 aagtccgatc cctcgcagga acagatcggc gctttcggcg ccttcgctgt ttttatcgcg
     1381 agcccagata atctgtttca tccgagcgca ctgaaataca tggcggacag aaatctcgcc
     1441 aaaatggaga tatggagaga gcaagctagt tgagttacct accactttct tggagttctt
     1501 cgcgtagtca atcagttgtt tttcgatgaa ctcgttaagc agcttgtcgg cattagacca
     1561 gccggggctc caggctctag tgagaagtgc attgcttggc ttttctgctt cattttccag
     1621 accgagttcc tcgatggagc aggcccagat ggcctctgca gcagcagtaa tcggcatcag
     1681 tctccaggga gggggcaaca taacggactc tattgacata tccaaacatt tcttccagta
     1741 actgttgaag gaggtgaacg gtttcccttt ctcgcaatag atctcccatg gctcgtacag
     1801 aaggtccccg ttgtagctct gcacactgat cccccgttcc accagcttct ccttcacagt
     1861 gtggtcgcgc accagactaa ctggatcgta caggtggtta aacacgactt tcgttgcgcc
     1921 ggtcaccctg atgcagtcga ggatggcaga gatggtattg tgtgtcttga tcagggtaag
     1981 atcactgccc agtgctttca gtgactggct aagatgggcc agactttgct tcatccacca
     2041 cctagaggcc cttccgggat aaaactggcc ttcttcttca ggacaccaaa tgaaaacggg
     2101 gaagacggaa ccctcgtggg ccgcggcggc gagcgcggga ttatcttcta ttctcaaatc
     2161 tctccgaaac cagacgatgg tctttttgtc catcttcata cctgtcggat ccgcgcctag
     2221 aatgtccagg tagacgggcg acgccttcgc caagttctga aggagtgtgt ggatgttctt
     2281 gatgttcttc cttgtgtgtg gttcccgctg ccagcatccc agcatcagct cgtacacctc
     2341 ctggggacac gtgcgaggcc gctgaaggac tctgccctgg gtgatgcatt ctatcacctc
     2401 gttgtttgat agctgatacc agggctgctt gccgtaggtg aagatctccc acaacacaac
     2461 tcccaggctc cagacgtcac tctcggtggt gaatttcctg tacatgatgc tctctggagg
     2521 catccatcgg atgggcaaca ttgtgtggcc accaacccgg tagtagtcgg tgctgtatac
     2581 atcccgggac atcccgaagt ccccaatttt caccagcagg ttctctccta ccaagcagtt
     2641 ccgggtggcc aggtctcggt gcacgaagtg ttgggatgcc aggtagacca tgcctgctgc
     2701 gatttgctga gcgatgtgca gcatctgcga ctgcgtcagc tcggtgggcg ggttaccctc
     2761 tgccatcagc actgcatctg gcccgtgtgc cctaaggaac ttgttgaggt ccccgtgctt
     2821 catgtactca aagaccatga tgagtgggtc gccctccaca cagacaccgt agaacttgac
     2881 aatgtgctcg tgctggaggt tggtcagcag ctcggcttcg cgatgaaagt ccttgcgagc
     2941 attgtcgctg gcgtccttca gcgtcttcac ggccaccagg atcttatcct gctcggggca
     3001 gaggttatag cactccgcta ggaaaacttt cccaaaggct ccttctccaa gctccctctt
     3061 cagaacgatg ttgtgtctct tgatgtgctg aacaaatgtg tccggcttga gctggctgtt
     3121 ggtgataccg aagtactggg ggttttcaat gacagggatc ttggtcatcc caatgatgac
     3181 agcatcgggc ccgccctccg aagaagacgg agtgttgctc ccgttggaga tgtggtggag
     3241 agggctggca gagtcatcgt cgttgctgat gacggaagct gggcctttca tgccaaactt
     3301 ggagtgtctt gccaacttgg aattcccagc actaccagca ctaccagcac tatctgctcc
     3361 cgcgctgtct ttcccttttg attttataca tcccatggtg gctagcaata acttcgtata
     3421 aagtatccta tacgaagtta tttgccttaa cccagaaatt atcactgtta ttctttagaa
     3481 tggtgcaaag aataacttcg tataatgtat gctatacgaa gttatgaatt cgatatcaag
     3541 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     3601 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     3661 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     3721 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     3781 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     3841 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     3901 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     3961 tggctgctcg cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     4021 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     4081 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     4141 cgataccgag cgctgctcga gagatctacg ggtggcatcc ctgtgacccc tccccagtgc
     4201 ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa
     4261 gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg
     4321 tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac
     4381 caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag
     4441 cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca
     4501 gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca
     4561 actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt
     4621 gaaccactgc tcccttccct gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg
     4681 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     4741 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     4801 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     4861 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     4921 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
     4981 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     5041 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     5101 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     5161 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     5221 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     5281 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     5341 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     5401 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     5461 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     5521 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     5581 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     5641 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     5701 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     5761 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     5821 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     5881 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     5941 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     6001 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     6061 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     6121 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     6181 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     6241 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     6301 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     6361 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     6421 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     6481 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     6541 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     6601 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     6661 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6721 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6781 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6841 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6901 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6961 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     7021 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     7081 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     7141 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     7201 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     7261 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     7321 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     7381 tttttacggt tcctggcctt ttgctggcct tttgctcaca tg
//
