LOCUS       sequence-433642-        7066 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 7066)
  AUTHORS   Huang Q, Chan KY, Wu J, Botticello-Romero NR, Zheng Q, Lou S, Keyes 
            C, Svanbergsson A, Johnston J, Mills A, Lin CY, Brauer PP, Clouse G,
            Pacouret S, Harvey JW, Beddow T, Hurley JK, Tobey IG, Powell M, Chen
            AT, Barry AJ, Eid FE, Chan YA, Deverman BE
  TITLE     An AAV capsid reprogrammed to bind human transferrin receptor 
            mediates brain-wide gene delivery.
  JOURNAL   Science. 2024 Jun 14;384(6701):1220-1227. doi: 
            10.1126/science.adm8386. Epub 2024 May 16.
  PUBMED    38753766
REFERENCE   2  (bases 1 to 7066)
  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"; doi: 
            "10.1126/science.adm8386"; journalName: "Science"; date: 
            "2024-06-14- 14"; volume: "384"; issue: "6701"; pages: "1220-1227"
COMMENT     Sequence Label: pAAV-CAG-SV40NLSf-mScarletP2A-hLuc
FEATURES             Location/Qualifiers
     source          1..7066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      143..598
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(230..249)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     440..461
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(615..634)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     734..756
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(794..812)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        880..984
                     /gene="bla"
                     /label=AmpR promoter
     promoter        985..1342
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        985..1176
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1008..1028)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1193..1328
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1377..2171
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1377..2171
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1431..1450)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2041..2060
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      2342..2930
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2831..2850
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2992..3121
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        3192..3495
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4104..4126
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     4188..4207
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             4295..4315
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4325..5017
                     /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(4455..4473)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4723..4742
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             5027..5083
                     /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"
     polyA_signal    6760..6881
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6771..6790)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6825..6844
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   join(6994..7066,1..68)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   join(6994..7066,1..57)
                     /label=AAV2 ITR
ORIGIN
        1 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
       61 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
      121 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
      181 gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct
      241 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
      301 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
      361 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
      421 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc
      481 tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat
      541 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
      601 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
      661 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
      721 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
      781 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
      841 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
      901 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
      961 caataatatt gaaaaaggaa gagtctgagg cggaaagaac cagctgtgga atgtgtgtca
     1021 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     1081 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     1141 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     1201 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     1261 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     1321 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     1381 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     1441 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     1501 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     1561 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     1621 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     1681 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     1741 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     1801 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagaac
     1861 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     1921 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     1981 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     2041 cgttggctac ccgtgatatt gctgaagaac ttggcggcga atgggctgac cgcttcctcg
     2101 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     2161 agttcttctg actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     2221 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     2281 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     2341 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     2401 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2461 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     2521 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2581 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     2641 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     2701 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     2761 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     2821 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     2881 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     2941 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg tcctgcaggc
     3001 agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt tggtcgcccg
     3061 gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc
     3121 tgcggccgca ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     3181 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     3241 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     3301 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     3361 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     3421 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     3481 atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc
     3541 ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg
     3601 gcgggggggg ggggggggcg cgcgccrggg gggggggggg ggggsggggg gsggggsggg
     3661 gsgrggcggr grggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt
     3721 atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc
     3781 gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg
     3841 ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc
     3901 tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg tgaaagcctt
     3961 gaggggctcc gggagcgccg gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc
     4021 gcgccgccgt ccccttctcc ctctccagcc tcggggctgt ccgcgggggg acggctgcct
     4081 tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gctctagagc
     4141 ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt
     4201 tattgtgctg tctcatcatt ttggcaaaga attgacgcgt gccaccatgt ctagtgatga
     4261 tgaggctact gctgactctc aacattctac tcctccaaaa aagaagagaa aggtagaaga
     4321 ccccgtgagc aagggcgagg cagtgatcaa ggagttcatg cggttcaagg tgcacatgga
     4381 gggctccatg aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga
     4441 gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct tctcctggga
     4501 catcctgtcc cctcagttca tgtacggctc cagggccttc accaagcacc ccgccgacat
     4561 ccccgactac tataagcagt ccttccccga gggcttcaag tgggagcgcg tgatgaactt
     4621 cgaggacggc ggcgccgtga ccgtgaccca ggacacctcc ctggaggacg gcaccctgat
     4681 ctacaaggtg aagctccgcg gcaccaactt ccctcctgac ggccccgtaa tgcagaagaa
     4741 gacaatgggc tgggaagcgt ccaccgagcg gttgtacccc gaggacggcg tgctgaaggg
     4801 cgacattaag atggccctgc gcctgaagga cggaggccgc tacctggcgg acttcaagac
     4861 cacctacaag gccaagaagc ccgtgcagat gcccggcgcc tacaacgtgg accgcaagtt
     4921 ggacatcacc tcccacaacg aggactacac cgtggtggaa cagtacgaac gctccgaggg
     4981 ccgccactcc accggcggca tggacgagct gtataaggga agcggagcta ctaacttcag
     5041 cctgctgaag caggctggag acgtggagga gaaccctggg cccgaggatg ccaagaatat
     5101 taagaaaggc cctgccccat tctaccctct ggaagatggc actgctggtg agcaactgca
     5161 caaggccatg aagaggtatg ccctggtccc tggcaccatt gccttcactg atgctcacat
     5221 tgaggtggac atcacctatg ctgaatactt tgagatgtct gtgaggctgg cagaagccat
     5281 gaaaagatat ggactgaaca ccaaccacag gattgtggtg tgctctgaga actctctcca
     5341 gttcttcatg cctgtgttag gagccctgtt cattggagtg gctgtggccc ctgccaatga
     5401 catctacaat gagagagagc tcctgaacag catgggcatc agccagccaa ctgtggtctt
     5461 tgtgagcaag aagggcctgc aaaagatcct gaatgtgcag aagaagctgc ccatcatcca
     5521 gaagatcatc atcatggaca gcaagactga ctaccagggc ttccagagca tgtatacctt
     5581 tgtgaccagc cacttacccc ctggcttcaa tgagtatgac tttgtgcctg agagctttga
     5641 cagggacaag accattgctc tgattatgaa cagctctggc tccactggac tgcccaaagg
     5701 tgtggctctg ccccacagaa ctgcttgtgt gagattcagc catgccagag accccatctt
     5761 tggcaaccag atcatccctg acactgccat cctgtctgtg gttccattcc atcatggctt
     5821 tggcatgttc acaacactgg ggtacctgat ctgtggcttc agagtggtgc tgatgtatag
     5881 gtttgaggag gagctgtttc tgaggagcct acaagactac aagatccagt ctgccctgct
     5941 ggtgcccact ctgttcagct tctttgccaa gagcaccctc attgacaagt atgacctgag
     6001 caacctgcat gagattgcct ctggaggagc acccctgagc aaggaggtgg gtgaggctgt
     6061 ggcaaagagg ttccatctcc caggaatcag acagggctat ggcctgactg agaccacctc
     6121 tgccatcctc atcacccctg aaggagatga caagcctggt gctgtgggca aggtggttcc
     6181 cttttttgag gccaaggtgg tggacctgga cactggcaag accctgggag tgaaccagag
     6241 gggtgagctg tgtgtgaggg gtcccatgat catgtctggc tatgtgaaca accctgaggc
     6301 caccaatgcc ctgattgaca aggatggctg gctgcactct ggtgacattg cctactggga
     6361 tgaggatgag cactttttca ttgtggacag gctgaagagc ctcatcaagt acaaaggcta
     6421 ccaagtggca cctgctgagc tagagagcat cctgctccag caccccaaca tctttgatgc
     6481 tggtgtggct ggcctgcctg atgatgatgc tggagagctg cctgctgctg ttgtggttct
     6541 ggagcatgga aagaccatga ctgagaagga gattgtggac tatgtggcca gtcaggtgac
     6601 cactgccaag aagctgaggg gaggtgtggt gtttgtggat gaggtgccaa agggtctgac
     6661 tggcaagctg gatgccagaa agatcagaga gatcctgatc aaggccaaga agggtggcaa
     6721 aatcgccgtc tagaattcct cgagttcgag cagacatgat aagatacatt gatgagtttg
     6781 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     6841 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     6901 attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag taaaacctct
     6961 acaaatgtgg taaaatccgg accgagcggc cgcaggaacc cctagtgatg gagttggcca
     7021 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaa
//
