LOCUS       sequence-402556-        7292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV reporter genome plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7292)
  AUTHORS   Huang Q, Chen AT, Chan KY, Sorensen H, Barry AJ, Azari B, Zheng Q, 
            Beddow T, Zhao B, Tobey IG, Moncada-Reid C, Eid FE, Walkey CJ, 
            Ljungberg MC, Lagor WR, Heaney JD, Chan YA, Deverman BE
  TITLE     Targeting AAV vectors to the central nervous system by engineering 
            capsid-receptor interactions that enable crossing of the blood-brain
            barrier.
  JOURNAL   PLoS Biol. 2023 Jul 19;21(7):e3002112. doi: 
            10.1371/journal.pbio.3002112. eCollection 2023 Jul.
  PUBMED    37467291
REFERENCE   2  (bases 1 to 7292)
  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: "PLoS 
            Biol."; date: "2023-07-19"; volume: "21(7):e3002112. doi"; pages: " 
            10.1371/journal.pbio.3002112. eCollection 2023 Jul"
COMMENT     Sequence Label: AAV-CAG-SV40NLSf-mScarlet-P2A-hLuc-WPRE
FEATURES             Location/Qualifiers
     source          1..7292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        201..504
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1073..1095
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1157..1176
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1264..1284
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1294..1986
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGGA
                     VTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKM
                     ALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHS
                     TGGMDELYK"
     primer_bind     complement(1424..1442)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1692..1711
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1996..2052
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     misc_feature    3709..4297
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3762..3782)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4180..4191)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4321..4442
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4332..4351)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4386..4405
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4555..4695
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4555..4684
                     /label=AAV2 ITR
     rep_origin      4770..5225
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4857..4876)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5067..5088
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5242..5261)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5361..5383
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5421..5439)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5507..5611
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5612..5680,5681..6472)
                     /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(5830..5849)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6643..7231
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7132..7151
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac tagttattaa tagtaatcaa ttacggggtc attagttcat
      181 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      241 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      301 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      361 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      421 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      481 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      541 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      601 gcgatggggg cggggggggg gggggsgggg cgaggcggag aggtgcggcg gcagccaatc
      661 agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata
      721 aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct
      781 ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga
      841 gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg
      901 tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagcgccgg caggaaggaa
      961 atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct
     1021 cggggctgtc cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc
     1081 ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt
     1141 ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa
     1201 ttgacgcgtg ccaccatgtc tagtgatgat gaggctactg ctgactctca acattctact
     1261 cctccaaaaa agaagagaaa ggtagaagac cccgtgagca agggcgaggc agtgatcaag
     1321 gagttcatgc ggttcaaggt gcacatggag ggctccatga acggccacga gttcgagatc
     1381 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
     1441 aagggtggcc ccctgccctt ctcctgggac atcctgtccc ctcagttcat gtacggctcc
     1501 agggccttca ccaagcaccc cgccgacatc cccgactact ataagcagtc cttccccgag
     1561 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgccgtgac cgtgacccag
     1621 gacacctccc tggaggacgg caccctgatc tacaaggtga agctccgcgg caccaacttc
     1681 cctcctgacg gccccgtaat gcagaagaag acaatgggct gggaagcgtc caccgagcgg
     1741 ttgtaccccg aggacggcgt gctgaagggc gacattaaga tggccctgcg cctgaaggac
     1801 ggaggccgct acctggcgga cttcaagacc acctacaagg ccaagaagcc cgtgcagatg
     1861 cccggcgcct acaacgtgga ccgcaagttg gacatcacct cccacaacga ggactacacc
     1921 gtggtggaac agtacgaacg ctccgagggc cgccactcca ccggcggcat ggacgagctg
     1981 tataagggaa gcggagctac taacttcagc ctgctgaagc aggctggaga cgtggaggag
     2041 aaccctgggc ccgaggatgc caagaatatt aagaaaggcc ctgccccatt ctaccctctg
     2101 gaagatggca ctgctggtga gcaactgcac aaggccatga agaggtatgc cctggtccct
     2161 ggcaccattg ccttcactga tgctcacatt gaggtggaca tcacctatgc tgaatacttt
     2221 gagatgtctg tgaggctggc agaagccatg aaaagatatg gactgaacac caaccacagg
     2281 attgtggtgt gctctgagaa ctctctccag ttcttcatgc ctgtgttagg agccctgttc
     2341 attggagtgg ctgtggcccc tgccaatgac atctacaatg agagagagct cctgaacagc
     2401 atgggcatca gccagccaac tgtggtcttt gtgagcaaga agggcctgca aaagatcctg
     2461 aatgtgcaga agaagctgcc catcatccag aagatcatca tcatggacag caagactgac
     2521 taccagggct tccagagcat gtataccttt gtgaccagcc acttaccccc tggcttcaat
     2581 gagtatgact ttgtgcctga gagctttgac agggacaaga ccattgctct gattatgaac
     2641 agctctggct ccactggact gcccaaaggt gtggctctgc cccacagaac tgcttgtgtg
     2701 agattcagcc atgccagaga ccccatcttt ggcaaccaga tcatccctga cactgccatc
     2761 ctgtctgtgg ttccattcca tcatggcttt ggcatgttca caacactggg gtacctgatc
     2821 tgtggcttca gagtggtgct gatgtatagg tttgaggagg agctgtttct gaggagccta
     2881 caagactaca agatccagtc tgccctgctg gtgcccactc tgttcagctt ctttgccaag
     2941 agcaccctca ttgacaagta tgacctgagc aacctgcatg agattgcctc tggaggagca
     3001 cccctgagca aggaggtggg tgaggctgtg gcaaagaggt tccatctccc aggaatcaga
     3061 cagggctatg gcctgactga gaccacctct gccatcctca tcacccctga aggagatgac
     3121 aagcctggtg ctgtgggcaa ggtggttccc ttttttgagg ccaaggtggt ggacctggac
     3181 actggcaaga ccctgggagt gaaccagagg ggtgagctgt gtgtgagggg tcccatgatc
     3241 atgtctggct atgtgaacaa ccctgaggcc accaatgccc tgattgacaa ggatggctgg
     3301 ctgcactctg gtgacattgc ctactgggat gaggatgagc actttttcat tgtggacagg
     3361 ctgaagagcc tcatcaagta caaaggctac caagtggcac ctgctgagct agagagcatc
     3421 ctgctccagc accccaacat ctttgatgct ggtgtggctg gcctgcctga tgatgatgct
     3481 ggagagctgc ctgctgctgt tgtggttctg gagcatggaa agaccatgac tgagaaggag
     3541 attgtggact atgtggccag tcaggtgacc actgccaaga agctgagggg aggtgtggtg
     3601 tttgtggatg aggtgccaaa gggtctgact ggcaagctgg atgccagaaa gatcagagag
     3661 atcctgatca aggccaagaa gggtggcaaa atcgccgtct agaattctaa tcaacctctg
     3721 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     3781 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     3841 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     3901 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     3961 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     4021 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     4081 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     4141 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     4201 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     4261 cagacgagtc ggatctccct ttgggccgcc tccccgcctc tcgagttcga gcagacatga
     4321 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     4381 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     4441 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
     4501 tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatccg gaccgagcgg ccgcaggaac
     4561 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     4621 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     4681 gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     4741 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
     4801 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc
     4861 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4921 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4981 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     5041 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     5101 ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt
     5161 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     5221 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     5281 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     5341 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     5401 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     5461 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5521 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     5581 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     5641 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     5701 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     5761 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     5821 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     5881 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     5941 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     6001 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     6061 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     6121 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     6181 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     6241 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6301 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     6361 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6421 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     6481 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6541 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6601 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6661 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6721 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6781 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6841 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6901 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6961 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     7021 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     7081 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     7141 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     7201 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     7261 cctggccttt tgctggcctt ttgctcacat gt
//
