LOCUS       sequence-442132-        5682 bp    DNA     circular SYN 15-APR-2025
DEFINITION  A recombinant AAV2 plasmid encoding vfChrimson-citrine with 
            trafficking enhancement..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5682)
  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 5682)
  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
FEATURES             Location/Qualifiers
     source          1..5682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(195..783)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(275..294)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(954..1745,1746..1814))
                     /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     1577..1596
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1815..1919)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1945..2400)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2082..2103)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2294..2313
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2472..2601
                     /label=AAV2 ITR
     promoter        2683..3130
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             join(4277..4279,4280..4282,4283..4993)
                     /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(4322..4343)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4583..4602)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4930..4951
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5064..5084)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   5511..5640
                     /label=AAV2 ITR
ORIGIN
        1 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
       61 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
      121 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
      181 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
      241 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
      301 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
      361 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
      421 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
      481 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
      541 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
      601 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
      661 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
      721 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
      781 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
      841 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
      901 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
      961 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     1021 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     1081 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     1141 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     1201 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     1261 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     1321 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     1381 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     1441 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     1501 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     1561 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     1621 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     1681 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     1741 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     1801 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     1861 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     1921 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
     1981 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     2041 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     2101 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     2161 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     2221 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     2281 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     2341 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     2401 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     2461 tattacgcca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
     2521 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
     2581 actccatcac taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacgt
     2641 agccatgctc taggaagatc tctgcagagg gccctgcgta tgagtgcaag tgggttttag
     2701 gaccaggatg aggcggggtg ggggtgccta cctgacgacc gaccccgacc cactggacaa
     2761 gcacccaacc cccattcccc aaattgcgca tcccctatca gagaggggga ggggaaacag
     2821 gatgcggcga ggcgcgtgcg cactgccagc ttcagcaccg cggacagtgc cttcgccccc
     2881 gcctggcggc gcgcgccacc gccgcctcag cactgaaggc gcgctgacgt cactcgccgg
     2941 tcccccgcaa actccccttc ccggccacct tggtcgcgtc cgcgccgccg ccggcccagc
     3001 cggaccgcac cacgcgaggc gcgagatagg ggggcacggg cgcgaccatc tgcgctgcgg
     3061 cgccggcgac tcagcgctgc ctcagtctgc ggtgggcagc ggaggagtcg tgtcgtgcct
     3121 gagagcgcag tcgaattcaa gcttggatcc gaccatggtg agcagaagac cctggctgct
     3181 ggccctggcc ctggccgtgg ccctggccgc cggcagcgcc ggcgccagca ccggcagcga
     3241 cgccaccgtg cccgtggcca cccaggacgg ccccgactac gtgttccaca gagcccacga
     3301 gagaatgctg ttccagacca gctacaccct ggagaacaac ggcagcgtga tctgcatccc
     3361 caacaacggc cagtgcttct gcctggcctg gctgctgcac agcagaggca caccaggaga
     3421 aaagatcggc gcccaggtct gccagtggat tgctttcagc atcgccatcg tcctgctgac
     3481 attctacggc ttcagcgcct ggaaggccac ttgcggttgg gaggaggtct acgtctgttg
     3541 cgtcgaggtg ctgttcgtga ccctggagat cttcaaggag ttcagcagcc ccgccacagt
     3601 gtacctgtct accggcaacc acgcctattg cctgcgctac ttcgagtggc tgctgtcttg
     3661 ccccgtgatc ctgatccgcc tgagcaacct gagcggcctg aagaacgact acagcaagcg
     3721 gaccatgggc ctgatcgtgt cttgcgtggg aatgatcgtg ttcggcatgg ccgcaggact
     3781 ggctaccgat tggctcaagt ggctgctgta tatcgtgtct tgcatctacg gcggctacat
     3841 gtacttccag gccgccaagt gctacgtgga agccaaccac agcgtgccta aaggccattg
     3901 ccgcatggtc gtgaagctga tggcctacgc tttcttcgcc tcttggggca tgtacccaat
     3961 cctctgggca gtgggaccag aaggactgct gaagctgagc ccttacgcca acagcatcgg
     4021 ccacagcatc tgcgacatca tcgccaagga gttttggacc ttcctggccc accacctgag
     4081 gatcaagatc cacgagcaca tcctgatcca cggcgacatc cggaagacca ccaagatgga
     4141 gatcggaggc gaggaggtgg aagtggaaga gttcgtggag gaggaggacg aggacacagt
     4201 ggcggccgcc aagagcagga tcaccagcga gggcgagtac atccccctgg accagatcga
     4261 catcaacgtg ctcgagatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     4321 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     4381 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     4441 cgtgccctgg cccaccctcg tgaccacctt cggctacggc ctgatgtgct tcgcccgcta
     4501 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     4561 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     4621 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     4681 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     4741 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     4801 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     4861 gctgcccgac aaccactacc tgagctacca gtccgccctg agcaaagacc ccaacgagaa
     4921 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     4981 cgagctgtac aagtagatcg attctagaat aatcaacctc tggattacaa aatttgtgaa
     5041 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     5101 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     5161 tcctggttag ttcttgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     5221 ggggctcggc tgttgggcac tgacaattcc gtggtgttta tttgtgaaat ttgtgatgct
     5281 attgctttat ttgtaaccat cactagttag gctttatttg tgaaatttgt gatgctattg
     5341 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     5401 ttatgtttca ggttcagggg gagatgtggg aggtttttta aaatcgataa ggatcttcct
     5461 agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct
     5521 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     5581 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     5641 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tg
//
