LOCUS       sequence-458355-        8397 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The mini version (8.4 kb) of Ad helper plasmid to provide E2A, 
            E4orf6, and VA RNA for AAV production.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8397)
  AUTHORS   Yang R, Tran NT, Chen T, Cui M, Wang Y, Sharma T, Liu Y, Zhang J, 
            Yuan X, Zhang D, Chen C, Shi Z, Wang L, Dai Y, Zaidi H, Liang J, 
            Chen M, Jaijyan D, Hu H, Wang B, Xu C, Hu W, Gao G, Yu D, Tai PWL, 
            Wang Q
  TITLE     AAVone: A cost-effective, single-plasmid solution for efficient AAV 
            production with reduced DNA impurities.
  JOURNAL   Mol Ther Nucleic Acids. 2025 May 14;36(2):102563. doi: 
            10.1016/j.omtn.2025.102563. eCollection 2025 Jun 10.
  PUBMED    40503174
REFERENCE   2  (bases 1 to 8397)
  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: "Mol Ther 
            Nucleic Acids."; date: "2025-05-14"; volume: "36(2):102563. doi"; 
            pages: " 10.1016/j.omtn.2025.102563. eCollection 2025 Jun 10"
COMMENT     Sequence Label: mini-pHelper-1.0
FEATURES             Location/Qualifiers
     source          1..8397
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      6133..6587
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6219..6238)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6429..6450
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6613..6717
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6718..7512
                     /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             6718..7512
                     /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(6772..6791)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7382..7401
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      7683..8271
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8172..8191
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atggtttaaa cgtcgacgta atccgtagat gtacctggac atccaggtga tgccggcggc
       61 ggtggtggag gcgcgcggaa agtcgcggac gcggttccag atgttgcgca gcggcaaaaa
      121 gtgctccatg gtcgggacgc tctggccggt gaggcgtgcg cagtcgttga cgctctagac
      181 cgtgcaaaag gagagcctgt aagcgggcac tcttccgtgg tctggtggat aaattcgcaa
      241 gggtatcatg gcggacgacc ggggttcgaa ccccggatcc ggccgtccgc cgtgatccat
      301 gcggttaccg cccgcgtgtc gaacccaggt gtgcgacgtc agacaacggg ggagcgctcc
      361 ttttggcttc cttccaggcg cggcggctgc tgcgctagct tttttggcca ctggccgcgc
      421 gcggcgtaag cggttaggct ggaaagcgaa agcattaagt ggctcgctcc ctgtagccgg
      481 agggttattt tccaagggtt gagtcgcagg acccccggtt cgagtctcgg gccggccgga
      541 ctgcggcgaa cgggggtttg cctccccgtc atgcaagacc ccgcttgcaa attcctccgg
      601 aaacagggac gagccccttt tttgcttttc ccagatgcat ccggtgctgc ggcagatgcg
      661 cccccctcct cagcagcggc aagagcaaga gcagcggcag acatgcaggg caccctcccc
      721 ttctcctacc gcgtcaggag gggcaacatc ctacatcgat tctagtgaat ccacagaaac
      781 tagcgaggta agcacttact ctatgtcttt tacatggtcc tgggaaagtg gaaaatacac
      841 cactgaaact tttgctacca actcttacac cttctcctac attgcccagg aataaagaat
      901 cgtgaacctg ttgcatgtta tgtttcaacg tgtttatttt tcaattgcag aaaatttcaa
      961 gtcatttttc attcagtagt atagccccac caccacatag cttatattga tcaccgtacc
     1021 ttaatcaaac tcacagaacc ctagtattca acctgccacc tccctcccaa cacacagagt
     1081 acacagtcct ttctccccgg ctggccttaa aaagcatcat atcatgggta acagacatat
     1141 tcttaggtgt tatattccac acggtttcct gtcgagccaa acgctcatca gtgatattaa
     1201 taaactcccc gggcagctcg cttaagttca tgtcgctgtc cagctgctga gccacaggct
     1261 gctgtccaac ttgcggttgc tcaacgggcg gcgaagggga agtccatgcc tacatggggg
     1321 tagagtcata atcgtgcatc aggatagggc ggtggtgctg cagcagcgcg cgaataaact
     1381 gctgccgccg ccgctccgtc ctgcaggaat acaacatggc agtggtctcc tcagcgatga
     1441 ttcgcaccgc ccgcagcatg agacgccttg tcctccgggc acagcagcgc accctgatct
     1501 cacttaaatc agcacagtaa ctgcagcaca gcaccacaat attgttcaaa atcccacagt
     1561 gcaaggcgct gtatccaaag ctcatggcgg ggaccacaga acccacgtgg ccatcatacc
     1621 acaagcgcag gtagattaag tggcgacccc tcataaacac gctggacata aacattacct
     1681 cttttggcat gttgtaattc accacctccc ggtaccatat aaacctctga ttaaacatgg
     1741 cgccatccac caccatccta aaccagctgg ccaaaacctg cccgccggct atgcactgca
     1801 gggaaccggg actggaacaa tgacagtgga gagcccagga ctcgtaacca tggatcatca
     1861 tgctcgtcat gatatcaatg ttggcacaac acaggcacac gtgcatacac ttcctcagga
     1921 ttacaagctc ctcccgcgtc agaaccatat cccagggaac aacccattcc tgaatcagcg
     1981 taaatcccac actgcaggga agacctcgca cgtaactcac gttgtgcatt gtcaaagtgt
     2041 tacattcggg cagcagcgga tgatcctcca gtatggtagc gcgggtctct gtctcaaaag
     2101 gaggtaggcg atccctactg tacggagtgc gccgagacaa ccgagatcgt gttggtcgta
     2161 gtgtcatgcc aaatggaacg ccggacgtag tcatatttcc tgaagcaaaa ccaggtgcgg
     2221 gcgtgacaaa cagatctgcg tctccggtct cgtcgcttag ctcgctctgt gtagtagttg
     2281 tagtatatcc actctctcaa agcatccagg cgcctaggca aaatagcacc ctcccgctcc
     2341 agaacaacat acagcgcttc cacagcggca gccataacag tcagccttac cagtaaaaaa
     2401 acctattaaa aaacaccact cgacacggca ccagctcaat cagtcacagt gtaaaaaggg
     2461 ccaagtacag agcgagtata tataggacta aaaaatgacg taacggttaa agtccacaaa
     2521 aaacacccag aaaaccgcac gcgaacctac gcccagaaac gaaagccaaa aaacccacaa
     2581 cttcctcaaa tcttcacttc cgttttccca cgatacgtca cttcccattt taaaaaaact
     2641 acaattccca atacatgcaa gttactccgc cctaaaacgc ccgggcgacc gcaccctgtg
     2701 acgaaagccg cccgcaagct gcgcccctga gttagtcatc tgaacttcgg cctgggcgtc
     2761 tctgggaagt accacagtgg tgggagcggg actttcctgg tacaccaggg cagcgggcca
     2821 actacgggga ttaaggttat tacgaggtgt ggtggtaata gccgcctgtt cgaggagaat
     2881 tcggtttcgg tgggcgcgga ttccgttgac ccgggatatc atgtggggtc ccgcgctcat
     2941 gtagtttatt cgggttgagt agtcttgggc agctccagcc gcaagtccca tttgtggctg
     3001 gtaactccac atgtagggcg tgggaatttc cttgctcata atggcgctga cgacaggtgc
     3061 tggcgccggg tgtggccgct ggagatgacg tagttttcgc gcttaaattt gagaaagggc
     3121 gcgaaactag tccttaagag tcagcgcgca gtatttgctg aagagagcct ccgcgtcttc
     3181 cagcgtgcgc cgaagctgat cttcgctttt gtgatacagg cagctgcggg tgagggagcg
     3241 cagagacctg ttttttattt tcagctcttg ttcttggccc ctgctttgtt gaaatatagc
     3301 atacagagtg ggaaaaatcc tatttctaag ctcgcgggtc gatacgggtt cgttgggcgc
     3361 cagacgcagc gctcctcctc ctgctgctgc cgccgctgtg gatttcttgg gctttgtcag
     3421 agtcttgcta tccggtcgcc tttgcttctg tgtgaccgct gctgttgctg ccgctgccgc
     3481 tgccgccggt gcagtagggg ctgtagagat gacggtagta atgcaggatg ttacggggga
     3541 aggccacgcc gtgatggtag agaagaaagc ggcgggcgaa ggagatgttg cccccacagt
     3601 cttgcaagca agcaactatg gcgttcttgt gcccgcgcca cgagcggtag ccttggcgct
     3661 gttgttgctc ttgggctaac ggcggcggct gcttagactt accggccctg gttccagtgg
     3721 tgtcccatct acggttgggt cggcgaacag gcagtgccgg cggcgcctga ggagcggagg
     3781 ttgtagcgat gctgggaacg gttgccaatt tctggggcgc cggcgagggg aatgcgaccg
     3841 agggtgacgg tgtttcgtct gacacctctt cggcctcgga agcttcgtct aggctgtccc
     3901 agtcttccat catctcctcc tcctcgtcca aaacctcctc tgcctgactg tcccagtatt
     3961 cctcctcgtc cgtgggtggc ggcggcggca gctgcagctt ctttttgggt gccatcctgg
     4021 gaagcaaggg cccgcggctg ctgatagggc tgcggcggcg gggggattgg gttgagctcc
     4081 tcgccggact gggggtccag gtaaaccccc cgtccctttc gtagcagaaa ctcttggcgg
     4141 gctttgttga tggcttgcaa ttggccaagg atgtggccct gggtaatgac gcaggcggta
     4201 agctccgcat ttggcgggcg ggattggtct tcgtagaacc taatctcgtg ggcgtggtag
     4261 tcctcaggga gaaggaaatg gccagtcggg aagaggagca gcgcgaaacc acccccgagc
     4321 gcggacgcgg tgcggcgcga cgtccaccaa ccatggagga cgtgtcgtcc ccgtcgccgt
     4381 cgccgccgcc tccccgcgcg cccccaaaaa agagactgcg gagacgcctg gagtccgagg
     4441 acgaggaaga cagttcccag gacgctctgg tgcctaggac accatctcca agaccctcca
     4501 cttccaccgc cgacctggcc attgccagca aaaaaaagaa gaagaggccc tcccctaaac
     4561 ctgaacgacc acccagccca gaagtgatcg tggactcaga ggaagagcgc gaggacgtcg
     4621 ctctgcagat ggtgggattc agcaatccac ctgtgctcat caagcacggc aaagggggaa
     4681 aacggaccgt gcgcaggctg aacgaggatg atcctgtggc aaggggcatg aggacacagg
     4741 aggagaagga agaatcctcc gaggccgaga gcgagagcac agtgattaat ccactgtccc
     4801 tgccaattgt gagcgcttgg gagaagggaa tggaggccgc ccgggccctg atggacaagt
     4861 accacgtgga caacgacctg aaggctaatt tcaaactgct gcctgaccag gtggaggccc
     4921 tggctgccgt gtgcaagaca tggctgaatg aggagcatag agggctgcag ctgaccttta
     4981 catcaaacaa gacctttgtg accatgatgg gcaggtttct gcaggcctac ctgcaatctt
     5041 tcgccgaagt gacttacaag caccacgagc ccaccggctg cgccctgtgg ctgcacagat
     5101 gcgccgagat tgagggcgag ctgaaatgcc tgcacggctc tatcatgatt aacaaggagc
     5161 acgtgatcga gatggatgtg acttccgaaa acggccagcg cgccctgaag gagcagtcca
     5221 gcaaggccaa gatcgtgaag aacaggtggg gcaggaacgt ggtgcagatt tctaacacag
     5281 acgccagatg ttgcgtgcat gacgccgcct gccctgccaa tcagtttagc ggcaagagct
     5341 gcggaatgtt cttcagcgag ggagccaagg cgcaagtggc cttcaagcag atcaaggcct
     5401 tcatgcaagc cctgtatcca aatgcccaga ccggccacgg ccacctcctg atgcccctgc
     5461 gctgcgaatg taactccaag cccggccacg cccctttcct gggcaggcag ctgccaaaac
     5521 tgaccccttt tgccctgagc aacgccgaag atctggatgc cgatctgatc agcgacaagt
     5581 ccgtgctggc ctccgttcac caccccgccc tgatcgtgtt ccagtgctgc aatcccgtgt
     5641 acaggaatag cagagctcag ggtggaggcc ccaactgcga cttcaagatc tccgccccag
     5701 atctgctgaa cgctctcgtg atggtgcgga gcctgtggtc tgagaacttc acagagctgc
     5761 cccgcatggt ggtgcccgag ttcaagtgga gcactaagca tcagtaccgg aatgtgtctc
     5821 tgcccgtggc ccactctgac gccagacaga atccttttga cttctgaacg gcgcagacgg
     5881 caagggtggg gggtaaataa tcacccgaga gtgtacaaat aaaaacattt gcctttattg
     5941 aaagtgtctc ctagtacatt atttttacat gtttttcaag tgacaaaaag aagtggcgct
     6001 cctaatctgc gcactgtggc tgcggaagta gggcgagtgg cgctccagga agctgtagag
     6061 ctgttcctgg ttgcgacgca gggtgggctg tacctgggga ctgttaagca tggagttggg
     6121 taccggatcc atcgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     6181 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     6241 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     6301 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     6361 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     6421 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     6481 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     6541 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     6601 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     6661 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     6721 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     6781 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     6841 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     6901 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtactc
     6961 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     7021 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     7081 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     7141 gagagagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     7201 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     7261 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     7321 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     7381 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     7441 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     7501 gagttcttct gactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     7561 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     7621 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     7681 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     7741 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     7801 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     7861 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     7921 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     7981 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     8041 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     8101 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     8161 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     8221 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     8281 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     8341 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgatcat
//
