LOCUS       sequence-486472-        5582 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV transfer plasmid expressing eGFP-MYC under a CAG promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5582)
  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 5582)
  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-MYC
FEATURES             Location/Qualifiers
     source          1..5582
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      346..934
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     835..854
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1171..1300
                     /label=AAV2 ITR
     polyA_signal    1462..1583
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1499..1518)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1553..1572
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    1626..2214
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1732..1743
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2141..2161
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2242..2271)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(2290..2319)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(2338..2367)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(join(2386..3096,3097..3099,3100..3102))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(2386..3096,3097..3099,3100..3102))
                     /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(2428..2449)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2777..2796
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3036..3057
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3099..3108
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3141..3160)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3222..3244)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     repeat_region   4110..4239
                     /label=AAV2 ITR
     rep_origin      4311..4766
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4398..4417)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4608..4629
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4792..4896
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4897..4965,4966..5582,1..175)
                     /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(5115..5134)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
       61 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
      121 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
      181 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
      241 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
      301 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
      361 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
      421 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
      481 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
      541 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
      601 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
      661 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
      721 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
      781 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
      841 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
      901 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
      961 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     1021 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     1081 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     1141 ccccgcgcgt tggccgattc attaatgcag ctgcgcgctc gctcgctcac tgaggccgcc
     1201 cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg
     1261 cgcagagagg gagtggccaa ctccatcact aggggttcct tgtagttaat gattaacccg
     1321 ccatgctact tatctacgta gccatgctct aggaagatcc ttatcgattt taccacattt
     1381 gtagaggttt tacttgcttt aaaaaacctc ccacatctcc ccctgaacct gaaacataaa
     1441 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     1501 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     1561 tccaaactca tcaatgtatc ttatcatgtc tgctcgaagc ggccgcccgg gtcgacggta
     1621 tcgatgcggg gaggcggccc aaagggagat ccgactcgtc tgagggcgaa ggcgaagacg
     1681 cggaagaggc cgcagagccg gcagcaggcc gcgggaagga aggtccgctg gattgagggc
     1741 cgaagggacg tagcagaagg acgtcccgcg cagaatccag gtggcaacac aggcgagcag
     1801 ccaaggaaag gacgatgatt tccccgacaa caccacggaa ttgtcagtgc ccaacagccg
     1861 agcccctgtc cagcagcggg caaggcaggc ggcgatgagt tccgccgtgg caatagggag
     1921 ggggaaagcg aaagtcccgg aaaggagctg acaggtggtg gcaatgcccc aaccagtggg
     1981 ggttgcgtca gcaaacacag tgcacaccac gccacgttgc ctgacaacgg gccacaactc
     2041 ctcataaaga gacagcaacc aggatttata caaggaggag aaaatgaaag ccatacggga
     2101 agcaatagca tgatacaaag gcattaaagc agcgtatcca catagcgtaa aaggagcaac
     2161 atagttaaga ataccagtca atctttcaca aattttgtaa tccagaggtt gattatcgat
     2221 aagcttgata tcgaattctt acaggtcctc ttcggagata agtttttgtt ctgatccacc
     2281 ggaacccccc aagtcttctt cgcttatcaa tttctgttcg ctacccccac tgccgcccag
     2341 gtcttcctct gaaataagtt tctgctcgct accgcctgag ccgcccttgt acagctcgtc
     2401 catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt
     2461 ctcgttgggg tctttgctca gggcggactg ggtgctcagg tagtggttgt cgggcagcag
     2521 cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc
     2581 gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg ccgttcttct gcttgtcggc
     2641 catgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc
     2701 gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa
     2761 cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg
     2821 gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta
     2881 gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc agggcacggg
     2941 cagcttgccg gtggtgcaaa tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc
     3001 gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct cgaccaggat
     3061 gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atggtggcgg atccaattct
     3121 ttgccaaaat gatgagacag cacaataacc agcacgttgc ccaggagctg taggaaaaag
     3181 aagaaggcat gaacatggtt agcagaggct ctagagccgc cggtcacacg ccagaagccg
     3241 aaccccgccc tgccccgtcc cccccgaagg cagccgtccc cccgcggaca gccccgagcc
     3301 gctccccccg cacaaagggc cctcccggag cccctcaagg ctttcacgca gccacagaaa
     3361 agaaacaagc cgtcattaaa ccaagcgcta attacagccc ggaggagaag ggccgtcccg
     3421 cccgctcacc tgtgggagta acgcggtcag tcagagccgg ggcgggcggc gcgaggcggc
     3481 ggcggagcgg ggcacggggc gaaggcagcg cgcagcgact cccgcccgcc gcgcgcttcg
     3541 ctttttatag ggccgccgcc gccgccgcct cgccataaaa ggaaactttc ggagcgcgcc
     3601 gctctgattg gctgccgccg cmcctcyccs ccycsccccs ccccsccccc cscccccccc
     3661 cccccccccy ggcgcgcgcc cccccccccc cccgccccca tcgctgcaca aaataattaa
     3721 aaaataaata aatacaaaat tgggggtggg gagggggggg agatggggag agtgaagcag
     3781 aacgtggggc tcacctcgac catggtaata gcgatgacta atacgtagat gtactgccaa
     3841 gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca tttaccgtca
     3901 ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc caagtgggca
     3961 gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg cgttactatg
     4021 ggaacatacg tcattattga cgtcaatggt actcttccta gagcatggct acgtagataa
     4081 gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt tggccactcc
     4141 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     4201 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tggcgtaata gcgaagaggc
     4261 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     4321 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4381 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4441 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4501 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     4561 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4621 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4681 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     4741 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     4801 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4861 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4921 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4981 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5041 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5101 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5161 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5221 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5281 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5341 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5401 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     5461 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5521 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5581 at
//
