LOCUS       sequence-442201-        5840 bp    DNA     circular SYN 15-APR-2025
DEFINITION  A recombinant AAV2 plasmid encoding vfChrimson-citrine with 
            trafficking enhancement and soma targeting with KV2.1 motif..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5840)
  AUTHORS   Mermet-Joret N, Moreno A, Zbela A, Ellendersen BE, Krauth N, von 
            Philipsborn A, Piriz J, Lin JY, Nabavi S
  TITLE     Dual-color optical activation and suppression of neurons with high 
            temporal precision
  JOURNAL   eLife 2023 12:RP90327
REFERENCE   2  (bases 1 to 5840)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "eLife 2023 
            12:RP90327"
COMMENT     Sequence Label: AAV-hSyn-IvfChr-citrine-KV2.1
FEATURES             Location/Qualifiers
     source          1..5840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     57..78
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(380..400)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     rep_origin      complement(1171..1759)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1251..1270)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1930..2721,2722..2790))
                     /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     2553..2572
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2791..2895)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2921..3376)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3058..3079)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3270..3289
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3448..3577
                     /label=AAV2 ITR
     promoter        3659..4106
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             join(5244..5246,5247..5249,5250..5840,1..120)
                     /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(5289..5310)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5550..5569)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 cactacctga gctaccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
       61 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
      121 cagtcccaac ccatactgaa cacgaaggaa atggctcctc aaagtaaacc gcccgaagaa
      181 ctcgaaatga gttcaatgcc atcacctgtg gctccgcttc ccgcccgcac ggaaggtgtc
      241 atcgacatgc gaagcatgtc tagtattgac tcatttatta gctgcgccac cgatttccct
      301 gaagctactc ggttctaatc tagaataatc aacctctgga ttacaaaatt tgtgaaagat
      361 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
      421 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
      481 ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
      541 ctcggctgtt gggcactgac aattccgtgg tgtttatttg tgaaatttgt gatgctattg
      601 ctttatttgt aaccatcact agttaggctt tatttgtgaa atttgtgatg ctattgcttt
      661 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
      721 gtttcaggtt cagggggaga tgtgggaggt tttttaaaat cgataaggat cttcctagag
      781 catggctacg tagataagta gcatggcggg ttaatcatta actacaagga acccctagtg
      841 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgc ccgggctttg
      901 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcat taatgaatcg gccaacgcgc
      961 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     1021 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     1081 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     1141 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     1201 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1261 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     1321 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     1381 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     1441 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     1501 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     1561 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     1621 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     1681 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     1741 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     1801 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     1861 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     1921 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     1981 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     2041 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     2101 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     2161 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     2221 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     2281 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     2341 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     2401 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     2461 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     2521 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     2581 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     2641 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     2701 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     2761 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     2821 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     2881 aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt
     2941 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     3001 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     3061 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     3121 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     3181 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     3241 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     3301 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     3361 ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg
     3421 gtgcgggcct cttcgctatt acgccagctg cgcgctcgct cgctcactga ggccgcccgg
     3481 gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc
     3541 agagagggag tggccaactc catcactagg ggttccttgt agttaatgat taacccgcca
     3601 tgctacttat ctacgtagcc atgctctagg aagatctctg cagagggccc tgcgtatgag
     3661 tgcaagtggg ttttaggacc aggatgaggc ggggtggggg tgcctacctg acgaccgacc
     3721 ccgacccact ggacaagcac ccaaccccca ttccccaaat tgcgcatccc ctatcagaga
     3781 gggggagggg aaacaggatg cggcgaggcg cgtgcgcact gccagcttca gcaccgcgga
     3841 cagtgccttc gcccccgcct ggcggcgcgc gccaccgccg cctcagcact gaaggcgcgc
     3901 tgacgtcact cgccggtccc ccgcaaactc cccttcccgg ccaccttggt cgcgtccgcg
     3961 ccgccgccgg cccagccgga ccgcaccacg cgaggcgcga gatagggggg cacgggcgcg
     4021 accatctgcg ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg ggcagcggag
     4081 gagtcgtgtc gtgcctgaga gcgcagtcga attcaagctt ggatccgacc atggtgagca
     4141 gaagaccctg gctgctggcc ctggccctgg ccgtggccct ggccgccggc agcgccggcg
     4201 ccagcaccgg cagcgacgcc accgtgcccg tggccaccca ggacggcccc gactacgtgt
     4261 tccacagagc ccacgagaga atgctgttcc agaccagcta caccctggag aacaacggca
     4321 gcgtgatctg catccccaac aacggccagt gcttctgcct ggcctggctg ctgcacagca
     4381 gaggcacacc aggagaaaag atcggcgccc aggtctgcca gtggattgct ttcagcatcg
     4441 ccatcgtcct gctgacattc tacggcttca gcgcctggaa ggccacttgc ggttgggagg
     4501 aggtctacgt ctgttgcgtc gaggtgctgt tcgtgaccct ggagatcttc aaggagttca
     4561 gcagccccgc cacagtgtac ctgtctaccg gcaaccacgc ctattgcctg cgctacttcg
     4621 agtggctgct gtcttgcccc gtgatcctga tccgcctgag caacctgagc ggcctgaaga
     4681 acgactacag caagcggacc atgggcctga tcgtgtcttg cgtgggaatg atcgtgttcg
     4741 gcatggccgc aggactggct accgattggc tcaagtggct gctgtatatc gtgtcttgca
     4801 tctacggcgg ctacatgtac ttccaggccg ccaagtgcta cgtggaagcc aaccacagcg
     4861 tgcctaaagg ccattgccgc atggtcgtga agctgatggc ctacgctttc ttcgcctctt
     4921 ggggcatgta cccaatcctc tgggcagtgg gaccagaagg actgctgaag ctgagccctt
     4981 acgccaacag catcggccac agcatctgcg acatcatcgc caaggagttt tggaccttcc
     5041 tggcccacca cctgaggatc aagatccacg agcacatcct gatccacggc gacatccgga
     5101 agaccaccaa gatggagatc ggaggcgagg aggtggaagt ggaagagttc gtggaggagg
     5161 aggacgagga cacagtgctc gagaagagca ggatcaccag cgagggcgag tacatccccc
     5221 tggaccagat cgacatcaac gtgatggtga gcaagggcga ggagctgttc accggggtgg
     5281 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     5341 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     5401 agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg atgtgcttcg
     5461 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     5521 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     5581 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     5641 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     5701 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     5761 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     5821 ccgtgctgct gcccgacaac
//
