LOCUS       sequence-485993-        5599 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV transfer plasmid expressing eGFP-Tag100 under a CAG promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5599)
  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 5599)
  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-Tag100
FEATURES             Location/Qualifiers
     source          1..5599
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(26..817,818..886))
                     /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     649..668
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(887..991)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1017..1472)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1154..1175)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1366..1385
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1544..1673
                     /label=AAV2 ITR
     promoter        1984..2259
                     /label=chicken beta-actin promoter
     primer_bind     2538..2560
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2622..2641
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      2674..2683
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2680..2682,2683..2685,2686..3396)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2680..2682,2683..2685,2686..3396)
                     /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(2725..2746)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2986..3005)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3333..3354
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3415..3450
                     /codon_start=1
                     /product="epitope tag derived from MAP kinase 2"
                     /label=Tag-100
                     /translation="EETARFQPGYRS"
     CDS             3469..3504
                     /codon_start=1
                     /product="epitope tag derived from MAP kinase 2"
                     /label=Tag-100
                     /translation="EETARFQPGYRS"
     CDS             3523..3558
                     /codon_start=1
                     /product="epitope tag derived from MAP kinase 2"
                     /label=Tag-100
                     /translation="EETARFQPGYRS"
     misc_feature    3586..4174
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3639..3659)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4057..4068)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4217..4338
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4228..4247)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4282..4301
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4500..4629
                     /label=AAV2 ITR
     rep_origin      complement(4866..5454)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4946..4965)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
       61 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
      121 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
      181 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
      241 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
      301 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
      361 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
      421 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
      481 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
      541 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
      601 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
      661 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
      721 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
      781 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
      841 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
      901 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
      961 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat
     1021 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     1081 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     1141 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     1201 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     1261 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     1321 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     1381 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     1441 cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg
     1501 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctgcgcg ctcgctcgct
     1561 cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt
     1621 gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt ccttgtagtt
     1681 aatgattaac ccgccatgct acttatctac gtagccatgc tctaggaaga gtaccattga
     1741 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     1801 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     1861 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     1921 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     1981 tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc
     2041 caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg
     2101 gggggcgcnn caggcggggc ggggcggggc grggggcggg gcggggcgag gcggagaggt
     2161 gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg
     2221 cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt
     2281 cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg
     2341 ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg
     2401 gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag
     2461 ggccctttgt gcggggggag cggctcgggg ctgtccgcgg ggggacggct gccttcgggg
     2521 gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc
     2581 taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc tggttattgt
     2641 gctgtctcat cattttggca aagaattgga tccgccacca tggtgagcaa gggcgaggag
     2701 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     2761 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     2821 atttgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     2881 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     2941 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     3001 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     3061 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     3121 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     3181 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     3241 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     3301 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     3361 gccgggatca ctctcggcat ggacgagctg tacaagggcg ggtccggagg aagcgaagaa
     3421 acagcacggt tccaaccagg gtaccggtca ggtggttccg gtgggtctga ggagacagcc
     3481 aggtttcaac ccggatatag atccggagga tctggaggct cagaagaaac tgccagattt
     3541 caacctggat ataggtccta agaattcgat atcaagctta tcgataatca acctctggat
     3601 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     3661 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     3721 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     3781 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     3841 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     3901 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     3961 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
     4021 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     4081 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     4141 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc cgggcggccg
     4201 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     4261 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     4321 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     4381 gagatgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag
     4441 gatcttccta gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa
     4501 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     4561 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     4621 agcgcgcagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4681 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4741 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4801 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4861 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4921 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4981 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5041 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5101 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5161 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5221 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5281 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5341 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5401 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5461 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5521 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5581 tttaaatcaa tctaaagta
//
