LOCUS       sequence-459855-        4808 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Single stranded AAV genome with concatemerization-dependent 
            barcodes, for tracking AAV concatemers via SpECTr. Also expresses 
            TagBFP from SCP1 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4808)
  AUTHORS   Coughlin GM, Borsos M, Barcelona BH, Appling N, Mayfield AMH, Mackey
            ED, Eser RA, Jackson CR, Chen X, Kumar SR, Gradinaru V
  TITLE     Spatial genomics of AAV vectors reveals mechanism of transcriptional
            crosstalk that enables targeted delivery of large genetic cargo.
  JOURNAL   Nat Biotechnol. 2025 Mar 20. doi: 10.1038/s41587-025-02565-4.
  PUBMED    40113953
REFERENCE   2  (bases 1 to 4808)
  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: "Nat 
            Biotechnol. 2025 Mar 20. doi: 10.1038/s41587-025-02565-4."
COMMENT     Sequence Label: pAAV.BC(p1-10)-SCP1-TagBFP2-W3SL-BC(p1-10)
FEATURES             Location/Qualifiers
     source          1..4808
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(5..27)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     127..146
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(163..618)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(300..321)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     512..531
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   693..833
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   704..833
                     /label=AAV2 ITR
     intron          1203..1335
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     CDS             1348..2049
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(2111..2131)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   2763..2903
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2763..2892
                     /label=AAV2 ITR
     rep_origin      complement(2965..3553)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3045..3064)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3724..4515,4516..4584))
                     /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     4347..4366
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4585..4689)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4757..4775
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
       61 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
      121 actatgcggc atcagagcag attgtactga gagtgcacca taaaattgta aacgttaata
      181 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caatagaccg
      241 aaatcggcaa aatcccttat aaatcaaaag aatagcccga gatagagttg agtgttgttc
      301 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
      361 ccgtctatca gggcgatggc ccactacgtg aaccatcacc caaatcaagt tttttggggt
      421 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
      481 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
      541 aggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
      601 cgccgctaca gggcgcgtac tatggttgct ttgacgtatg cggtgtgaaa taccgcacag
      661 atgcgtaagg agaaaatacc gcatcaggcg cccctgcagg cagctgcgcg ctcgctcgct
      721 cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt
      781 gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt cctgctgaca
      841 ggaggattct ctcatcactg aaacaaaaga agctgttcca atccacgaac ccgaacctga
      901 atcactttct ttgaattttt tgcgactggt gttcaagctg aatttaagtt cccggtatta
      961 gacaaattcc tggccatgcc ttttatttta gaatgacgac atttccgcct tgtaccttat
     1021 tctgggttgt ttcacaagtg gggttattac accatgtgat atgccaacgt caattcttgc
     1081 tgtgtacctg catggtaatg tgatgggccg cctcgaggta cttatataag ggggtggggg
     1141 cgcgttcgtc ctcagtcgcg atcgaacact cgagccgagc agacgtgcct acggaccgac
     1201 aggtaagtat caaggttaca agacaggttt aaggagacca atagaaactg ggcttgtcga
     1261 gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac atccactttg
     1321 cctttctctc cacagaccgg tgccaccatg agcgagctga ttaaggagaa catgcacatg
     1381 aagctgtaca tggagggcac cgtggacaac catcacttca agtgcacatc cgagggcgaa
     1441 ggcaagccct acgagggcac ccagaccatg agaatcaagg tggtcgaggg cggccctctc
     1501 cccttcgcct tcgacatcct ggctactagc ttcctctacg gcagcaagac cttcatcaac
     1561 cacacccagg gcatccccga cttcttcaag cagtccttcc ctgagggctt cacatgggag
     1621 agagtcacca catacgaaga cgggggcgtg ctgaccgcta cccaggacac cagcctccag
     1681 gacggctgcc tcatctacaa cgtcaagatc agaggggtga acttcacatc caacggccct
     1741 gtgatgcaga agaaaacact cggctgggag gccttcaccg agacgctgta ccccgctgac
     1801 ggcggcctgg aaggcagaaa cgacatggcc ctgaagctcg tgggcgggag ccatctgatc
     1861 gcaaacgcca agaccacata tagatccaag aaacccgcta agaacctcaa gatgcctggc
     1921 gtctactatg tggactacag actggaaaga atcaaggagg ccaacaacga gacctacgtc
     1981 gagcagcacg aggtggcagt ggccagatac tgcgacctcc ctagcaaact ggggcacaag
     2041 cttaattagg aattcataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     2101 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     2161 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttagttc
     2221 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     2281 tgggcactga caattccgtg gtgtttattt gtgaaatttg tgatgctatt gctttatttg
     2341 taaccatcta gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     2401 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     2461 gagatgtggg aggtttttta aacaaggtac catcacatta ccatgcaggt acacagcaag
     2521 aattgacgtt ggcatatcac atggtgtaat aaccccactt gtgaaacaac ccagaataag
     2581 gtacaaggcg gaaatgtcgt cattctaaaa taaaaggcat ggccaggaat ttgtctaata
     2641 ccgggaactt aaattcagct tgaacaccag tcgcaaaaaa ttcaaagaaa gtgattcagg
     2701 ttcgggttcg tggattggaa cagcttcttt tgtttcagtg atgagagaat cctcctgtca
     2761 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     2821 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgagcggcct cagtgagcga
     2881 gcgagcgcgc agctgcctgc aggacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2941 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3001 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3061 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3121 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3181 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3241 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3301 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3361 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3421 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3481 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3541 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3601 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3661 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     3721 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     3781 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     3841 ccccagtgct gcaatgatac cgcggcttcc acgctcaccg gctccagatt tatcagcaat
     3901 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     3961 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4021 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4081 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4141 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4201 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4261 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4321 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4381 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4441 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4501 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4561 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     4621 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4681 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     4741 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     4801 tgacggtg
//
