LOCUS       sequence-379359-        7300 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV production for expression of optically controlled eDrTrkB with 
            N-terminal fusion of mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7300)
  AUTHORS   Leopold AV, Thankachan S, Yang C, Gerashchenko D, Verkhusha VV
  TITLE     A general approach for engineering RTKs optically controlled with 
            far-red light.
  JOURNAL   Nat Methods. 2022 Jul;19(7):871-880. doi: 
            10.1038/s41592-022-01517-z. Epub 2022 Jun 9.
  PUBMED    35681062
REFERENCE   2  (bases 1 to 7300)
  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: 
            "10.1038/s41592-022-01517-z"; journalName: "Nat Methods"; date: 
            "2022-07"; volume: "19"; issue: "7"; pages: "871-880"
COMMENT     Sequence Label: AAV2/9-CW3SL-mCherry-DrTrkB
FEATURES             Location/Qualifiers
     source          1..7300
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     primer_bind     538..557
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        538..556
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        562..580
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     562..579
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     sig_peptide     606..665
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             669..1370
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="SKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGTQ
                     TAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFED
                     GGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGE
                     IKQRLKLKDGCHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAEG
                     RHSTGGMDELYK"
     primer_bind     complement(808..826)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1076..1095
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(4183..4203)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4563..4703
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4563..4692
                     /label=AAV2 ITR
     rep_origin      4778..5233
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4865..4884)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5075..5096
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5250..5269)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5369..5391
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5429..5447)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5515..5619
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5620..5688,5689..6480)
                     /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(5838..5857)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6651..7239
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7140..7159
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcag atgctgcaca cttgtggact aagtttgttc gcatcccctt
      181 ctccaacccc ctcagtacat caccctgggg gaacagggtc cacttgctcc tgggcccaca
      241 cagtcctgca gtattgtgta tataaggcca gggcaaagag gagcaggttt taaagtgaaa
      301 ggcaggcagg tgttggggag gcagttaccg gggcaacggg aacagggcgt ttcggaggtg
      361 gttgccatgg ggacctggat gctgacgaag gctcgcgagg ctgtgagcag ccacagtgcc
      421 ctgctcagaa gccccaagct cgtcagtcaa gccggttctc cgtttgcact caggagcacg
      481 ggcaggcgag tggcccctag ttctgggggc agcgtgaaaa gacggtatcg ataagcttaa
      541 tacgactcac tatagggtac gatttaggtg acactataga ataaggagcg atcgccatgg
      601 ctagcatgga gacagacaca ctcctgctat gggtactgct gctctgggtt ccaggttcca
      661 ctggtgacag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
      721 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
      781 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
      841 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
      901 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
      961 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
     1021 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     1081 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     1141 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggctgc cactacgacg
     1201 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     1261 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     1321 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag gaattcagtg
     1381 ctggtagtgc tggtagtgct ggcaccggta tgagccggga cccgttgccc ttttttccac
     1441 cgctttacct tggtggcccg gaaattacca ccgagaactg cgagcgcgag ccgattcata
     1501 ttcccggcag catccagccg cacggcgccc tgctcactgc cgacgggcac agcggcgagg
     1561 tgctccagat gagcctcaac gcggccactt ttctgggaca ggaacccaca gtgctgcgcg
     1621 gacagaccct cgccgcactg ctgcccgagc agtggcccgc gctgcaagcg gccctgcccc
     1681 ccggctgccc cgacgccctg caataccgcg caacgctgga ctggcctgcc gccgggcacc
     1741 tttcgctgac ggtgcaccgg gtcggcgagt tgctgattct ggagttcgag ccgacggagg
     1801 cctgggacag caccgggccg cacgcgctgc gcaacgcgat gttcgcgctc gaaagtgccc
     1861 ccaacctgcg ggcgctggcc gaggtggcga cccagacggt ccgcgagctg acgggctttg
     1921 accgggtgat gctctacaaa tttgcccccg acgccaccgg cgaagtgatt gccgaggccc
     1981 gccgtgaggg gctgcacgcc tttctgggcc accgttttcc cgcgtcggac attccggcgc
     2041 aggcccgcgc gctctacacc cggcacctgc tgcgcctgac cgccgacacc cgcgccgccg
     2101 ccgtgccgct cgatcccgtc ctcaacccgc agacgaatgc gcccaccccg ctgggcggcg
     2161 ccgtgctgcg cgccacctcg cccatgcaca tgcagtacct gcggaacatg ggcgtcgggt
     2221 cgagcctgtc ggtgtcggtg gtggtcggcg gccagctctg gggcctgatc gcctgccacc
     2281 accagacgcc ctacgtgttg ccgcccgacc tgcgaaccac gctcgaatac ctgggccgct
     2341 tgctgagcct gcaagttcag gtcaaggaag cggcggacgt ggcggccttt cgccagagcc
     2401 tgcgggagca ccacgcgcgg gtggccctcg cggcggcgca ctcgctctcg ccgcacgaca
     2461 ccctcagtga cccggcgctt gacctgctgg gcctgatgcg ggccgggggc ctgattctgc
     2521 gtttcgaggg ccgctggcag acgttgggtg aagtgccgcc tgccccggcg gtggacgcgc
     2581 tgctggcgtg gctcgaaacc cagccgggcg ccctggtcca gaccgacgcg ctaggccaac
     2641 tgtggcccgc cggcgccgat ctcgccccca gcgcagcggg cctgctcgcc atcagcgtgg
     2701 gcgagggctg gtcggagtgc ctcgtctggc tgcggcccga actgcggctg gaggtcgcct
     2761 ggggcggggc cactcctgac caggcgaaag acgacctcgg gccgcgccac tcattcgaca
     2821 cctacctcga agaaaaacgc ggctacgccg agccctggca tcccggcgaa atcgaggagg
     2881 cgcaggatct acgtgacaca ttgaccgggg cgctgggcga gcagctcgag atcgccactg
     2941 ggatggtggg ggccctcctc ttgctgctgg tggtggccct ggggatcggc ctcttcatgt
     3001 ctagaaagtt ggcaagacac tccaagtttg gcatgaaagg cccagcttcc gtcatcagca
     3061 acgacgatga ctctgccagc cctctccacc acatctccaa cgggagcaac actccgtctt
     3121 cttcggaggg cgggcccgat gctgtcatca ttgggatgac caagatccct gtcattgaaa
     3181 acccccagta cttcggtatc accaacagcc agctcaagcc ggacacattt gttcagcaca
     3241 tcaagagaca caacatcgtt ctgaagaggg agcttggaga aggagccttt gggaaagttt
     3301 tcctagcgga gtgctataac ctctgccccg agcaggataa gatcctggtg gccgtgaaga
     3361 cgctgaagga cgccagcgac aatgctcgca aggactttca tcgcgaagcc gagctgctga
     3421 ccaacctcca gcacgagcac attgtcaagt tctacggtgt ctgtgtggag ggcgacccac
     3481 tcatcatggt ctttgagtac atgaagcacg gggacctcaa caagttcctt agggcacacg
     3541 ggccagatgc agtgctgatg gcagagggta acccgcccac cgagctgacg cagtcgcaga
     3601 tgctgcacat cgctcagcaa atcgcagcag gcatggtcta cctggcatcc caacacttcg
     3661 tgcaccgaga cctggccacc cggaactgct tggtaggaga gaacctgctg gtgaaaattg
     3721 gggacttcgg gatgtcccgg gatgtataca gcaccgacta ctaccgggtt ggtggccaca
     3781 caatgttgcc catccgatgg atgcctccag agagcatcat gtacaggaaa ttcaccaccg
     3841 agagtgacgt ctggagcctg ggagttgtgt tgtgggagat cttcacctac ggcaagcagc
     3901 cctggtatca gctatcaaac aacgaggtga tagaatgcat cacccagggc agagtccttc
     3961 agcggcctcg cacgtgtccc caggaggtgt acgagctgat gctgggatgc tggcagcggg
     4021 aaccacacac aaggaagaac atcaagaaca tccacacact ccttcagaac ttggcgaagg
     4081 cgtcgcccgt ctacctggac atcctaggct agtgatctag agtcgagata atcaacctct
     4141 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     4201 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     4261 tttctcctcc ttgtataaat cctggttagt tcttgccacg gcggaactca tcgccgcctg
     4321 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgtttat
     4381 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatc tagctttatt tgtgaaattt
     4441 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     4501 attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagcggcc
     4561 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     4621 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     4681 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     4741 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     4801 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
     4861 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4921 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4981 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     5041 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     5101 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc
     5161 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     5221 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     5281 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     5341 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     5401 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     5461 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     5521 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5581 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5641 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5701 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5761 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     5821 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     5881 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5941 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     6001 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     6061 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     6121 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     6181 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     6241 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6301 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     6361 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     6421 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6481 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6541 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6601 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6661 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6721 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6781 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6841 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6901 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6961 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     7021 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     7081 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     7141 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     7201 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     7261 ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
