LOCUS       sequence-486479-        5591 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV transfer plasmid expressing eGFP-VSV under a CAG promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5591)
  AUTHORS   Park SY, Sheridan A, An B, Jarvis E, Lyudchik J, Patton W, Axup JY, 
            Chan SW, Damstra HGJ, Leible D, Leung KS, Magno CA, Meeran A, 
            Michalska JM, Rieger F, Wang C, Wu M, Church GM, Funke J, Huffman T,
            Leeper KGC, Truckenbrodt S, Winnubst J, Kornfeld JMR, Boyden ES, 
            Rodriques SG, Payne AC
  TITLE     Combinatorial protein barcodes enable self-correcting neuron tracing
            with nanoscale molecular context
  JOURNAL   bioRxiv 2025.09.26.678648
REFERENCE   2  (bases 1 to 5591)
  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"; journalName: "bioRxiv 
            2025.09.26.678648"
COMMENT     Sequence Label: pAAV-CAG-eGFP-VSV
FEATURES             Location/Qualifiers
     source          1..5591
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     22..41
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(260..364)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(390..845)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(527..548)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     739..758
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   917..1046
                     /label=AAV2 ITR
     primer_bind     1912..1934
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1996..2015
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      2048..2057
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2054..2056,2057..2059,2060..2770)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2054..2056,2057..2059,2060..2770)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2099..2120)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2360..2379)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2707..2728
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2789..2821
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     CDS             2840..2872
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     CDS             2891..2923
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     misc_feature    2951..3539
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3004..3024)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3422..3433)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3582..3703
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3593..3612)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3647..3666
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3865..3994
                     /label=AAV2 ITR
     rep_origin      complement(4231..4819)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4311..4330)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4990..5591,1..190,191..259))
                     /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"
ORIGIN
        1 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
       61 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
      121 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
      181 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
      241 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
      301 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
      361 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt
      421 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
      481 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
      541 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
      601 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
      661 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
      721 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
      781 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
      841 cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
      901 ttcgctatta cgccagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg
      961 gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt
     1021 ggccaactcc atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc
     1081 tacgtagcca tgctctagga agagtaccat tgacgtcaat aatgacgtat gttcccatag
     1141 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     1201 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     1261 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     1321 agtacatcta cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct
     1381 tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt attttttaat
     1441 tattttgtgc agcgatgggg gcgggggggg ggggggggcg cgcscrgggg gggggggggg
     1501 gggggggggg gggggsgggg sgrggsggrg rggkgcggcg gcagccaatc agagcggcgc
     1561 gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag
     1621 cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc
     1681 ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga
     1741 cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct
     1801 gtggctgcgt gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc
     1861 ggggctgtcc gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct
     1921 tctggcgtgt gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt
     1981 tcctacagct cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat
     2041 tggatccgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     2101 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     2161 cgatgccacc tacggcaagc tgaccctgaa gttcatttgc accaccggca agctgcccgt
     2221 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     2281 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     2341 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     2401 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     2461 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     2521 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     2581 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     2641 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     2701 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     2761 gctgtacaag gggggtagcg gcggttccta cacagacata gagatgaacc gactcggtaa
     2821 aggtggcagt gggggctcat atactgatat tgaaatgaat agactcggga aggggggtag
     2881 tggcggcagt tataccgaca tagaaatgaa taggctggga aagtaagaat tcgatatcaa
     2941 gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     3001 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     3061 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     3121 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     3181 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     3241 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     3301 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     3361 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     3421 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     3481 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca
     3541 tcgataccgt cgacccgggc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     3601 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3661 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3721 tcattttatg tttcaggttc agggggagat gtgggaggtt ttttaaagca agtaaaacct
     3781 ctacaaatgt ggtaaaatcg ataaggatct tcctagagca tggctacgta gataagtagc
     3841 atggcgggtt aatcattaac tacaaggaac ccctagtgat ggagttggcc actccctctc
     3901 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     3961 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcat taatgaatcg gccaacgcgc
     4021 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4081 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4141 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4201 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4261 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4321 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4381 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4441 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4501 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4561 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4621 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4681 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4741 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4801 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4861 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4921 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4981 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5041 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5101 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5161 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5221 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5281 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5341 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5401 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5461 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5521 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5581 accgagttgc t
//
