LOCUS       sequence-455008-        8263 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Adenoviral helper plasmid containing E2A, VA RNA I and L4-22k genes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8263)
  AUTHORS   Doshi J, Couto E, Staiti J, Vandenberghe LH, Zabaleta N
  TITLE     E2A, VA RNA I, and L4-22k adenoviral helper genes are sufficient for
            AAV production in HEK293 cells.
  JOURNAL   Mol Ther Methods Clin Dev. 2024 Nov 13;32(4):101376. doi: 
            10.1016/j.omtm.2024.101376. eCollection 2024 Dec 12.
  PUBMED    39670178
REFERENCE   2  (bases 1 to 8263)
  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 
            Methods Clin Dev."; date: "2024-11-13"; volume: "32(4):101376. doi";
            pages: " 10.1016/j.omtm.2024.101376. eCollection 2024 Dec 12"
COMMENT     Sequence Label: JD226
FEATURES             Location/Qualifiers
     source          1..8263
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    52..123
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(128..174)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    192..240
                     /note="synthetic polyadenylation signal"
     promoter        277..1455
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          508..1446
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1403..1423
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1661..1670
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2301..2422
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2312..2331)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2366..2385
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    2431..2655
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(4411..4614)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4440..4460)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4615..4918
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     terminator      5266..5312
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      5342..5478
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5523..5594
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(5820..6408)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5900..5919)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(7154..7222,7223..8014)
                     /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(7372..7391)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      8027..8163
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg acatcttgga
       61 ccattagctc cacaggtatc ttcttccctc tagtggtcat aacagcagct tcagctacct
      121 ctcaattcaa aaaacccctc aagacccgtt tagaggcccc aaggggttat gctatcaatc
      181 gttgcgttac acacacaaaa aaccaacaca catccatctt cgatggatag cgattttatt
      241 atctaactgc tgatcgagtg tagccagatc tcgtgaggct ccggtgcccg tcagtgggca
      301 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt
      361 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt
      421 tttcccgagg gtgggggaga accgtatata agtgcactag tcgccgtgaa cgttcttttt
      481 cgcaacgggt ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc
      541 ctctttacgg gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg
      601 attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgtggcc ttgcgcttaa
      661 ggagcccctt cgcctcgtgc ttgagttgtg gcctggcctg ggcgctgggg ccgccgcgtg
      721 cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa
      781 aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc
      841 caagatcagc acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg
      901 tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg
      961 gggtagtctc aagctgcccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc
     1021 ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg
     1081 cttcccggcc ctgctgcagg gagcacaaaa tggaggacgc ggcgctcggg agagcgggcg
     1141 ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac
     1201 tccacggagt accgggcgcc gtccaggcac ctcgattagt tctccagctt ttggagtacg
     1261 tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg
     1321 gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt
     1381 gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca
     1441 tttcaggtgt cgtgaaaact acccctaaaa gccaaaagat ctttgtcgat cctaccatcc
     1501 actcgacaca cccgccagcg gccgctgcca agcttccgag ctctcgaatt catcttggct
     1561 gaggaatctt ctaacaattt agagcttaaa aacgcccacg aggcggagaa cgaaatatcc
     1621 agagagacgt tagaaacgtt caaaaacgtt caccggtagg gccaccatgg cacccaaaaa
     1681 gaagctgcag ctgccgccgc cacccacgga cgaggaggaa tactgggaca gtcaggcaga
     1741 ggaggttttg gacgaggagg aggaggacat gatggaagac tgggagagcc tagacgagga
     1801 agcttccgag gtcgaagagg tgtcagacga aacaccgtca ccctcggtcg cattcccctc
     1861 gccggcgccc cagaaatcgg caaccggttc cagcatggct acaacctccg ctcctcaggc
     1921 gccgccggca ctgcccgttc gccgacccaa ccgtagatgg gacaccactg gaaccagggc
     1981 cggtaagtcc aagcagccgc cgccgttagc ccaagagcaa caacagcgcc aaggctaccg
     2041 ctcatggcgc gggcacaaga acgccatagt tgcttgcttg caagactgtg ggggcaacat
     2101 ctccttcgcc cgccgctttc ttctctacca tcacggcgtg gccttccccc gtaacatcct
     2161 gcattactac cgtcatctct acagcccata ctgcaccggc ggcagcggca gcaacagcag
     2221 cggccacaca gaagcaaagg cgaccggata ggctagcttg actgactgag atacagcgta
     2281 ccttcagctc acagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     2341 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     2401 tataagctgc aataaacaag ttaggaatgg ccatagagcc caccgcatcc ccagcatgcc
     2461 tgctattgtg ttcccaatcc tcccccttgc tgtcctgccc caccccaccc cccagaatag
     2521 aatgacacct actcagacaa tgcgatgcaa tttcctcatt ttattaggaa aggacagtgg
     2581 gagtggcacc ttccagggtc aaggaaggca cgggggaggg gcaaacaaca gatggctggc
     2641 aactagaagg cacagtaaaa tagataaatg gcatcttgcc gtctgcgccg tttaaaaatc
     2701 aaaggggttc tgccgcgcat cgctatgcgc cactggcagg gacacgttgc gatactggtg
     2761 tttagtgctc cacttaaact caggcacaac catccgcggc agctcggtga agttttcact
     2821 ccacaggctg cgcaccatca ccaacgcgtt tagcaggtcg ggcgccgata tcttgaagtc
     2881 gcagttgggg cctccgccct gcgcgcgcga gttgcgatac acagggttgc agcactggaa
     2941 cactatcagc gccgggtggt gcacgctggc cagcacgctc ttgtcggaga tcagatccgc
     3001 gtccaggtcc tccgcgttgc tcagggcgaa cggagtcaac tttggtagct gccttcccaa
     3061 aaagggtgca tgcccaggct ttgagttgca ctcgcaccgt agtggcatca gaaggtgacc
     3121 gtgcccggtc tgggcgttag gatacagcgc ctgcatgaaa gccttgatct gcttaaaagc
     3181 cacctgagcc tttgcgcctt cagagaagaa catgccgcaa gacttgccgg aaaactgatt
     3241 ggccggacag gccgcgtcat gcacgcagca ccttgcgtcg gtgttggaga tctgcaccac
     3301 atttcggccc caccggttct tcacgatctt ggccttgcta gactgctcct tcagcgcgcg
     3361 ctgcccgttt tcgctcgtca catccatttc aatcacgtgc tccttattta tcataatgct
     3421 cccgtgtaga cacttaagct cgccttcgat ctcagcgcag cggtgcagcc acaacgcgca
     3481 gcccgtgggc tcgtggtgct tgtaggttac ctctgcaaac gactgcaggt acgcctgcag
     3541 gaatcgcccc atcatcgtca caaaggtctt gttgctggtg aaggtcagct gcaacccgcg
     3601 gtgctcctcg tttagccagg tcttgcatac ggccgccaga gcttccactt ggtcaggcag
     3661 tagcttgaag tttgccttta gatcgttatc cacgtggtac ttgtccatca acgcgcgcgc
     3721 agcctccatg cccttctccc acgcagacac gatcggcagg ctcagcgggt ttatcaccgt
     3781 gctttcactt tccgcttcac tggactcttc cttttcctct tgcgtccgca taccccgcgc
     3841 cactgggtcg tcctcattca gccgccgcac cgtgcgctta cctcccttgc cgtgcttgat
     3901 tagcaccggt gggttgctga aacccaccat ttgtagcgcc acatcttctc tttcttcctc
     3961 gctgtccacg atcacctctg gggatggcgg gcgctcgggc ttgggagagg ggcgcttctt
     4021 tttctttttg gacgcaatgg ccaaatccgc cgtcgaggtc gatggccgcg ggctgggtgt
     4081 gcgcggcacc agcgcatctt gtgacgagtc ctcttcgtcc tcggactcga gccgccgcct
     4141 cagccgcttt tttgggggcg cgcggggagg cggcggcgac ggcgacgggg acgacacgtc
     4201 ctccatggtt ggtggacgtc gcgccgcacc gcgtccgcgc tcgggggtgg tttcgcgctg
     4261 ctcctcttcc cgactggcca tgctggcggt accttccttc tcctgatatc gcctcgctcg
     4321 acgaagtgcc aaaaatcttt gagggtcttg gacgcgacga gaaacgcgcg gcaaacgctc
     4381 cgcagctgcc tgtatcacaa cctaggtctg agctctgctt atatagacct cccaccgtac
     4441 acgcctaccg cccatttgcg tcaatggggc ggagttgtta cgacattttg gaaagtcccg
     4501 ttgattttgg tgccaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc
     4561 ccgtgagtca aaccgctatc cacgcccatt gatgtactgc caaaaccgca tcaccatggt
     4621 aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta
     4681 ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg
     4741 catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg
     4801 acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa
     4861 tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgct
     4921 ccgtaccagc tccatggtcg ggacgctctg gccggtgagg cgtgcgcagt cgttgacgct
     4981 ctagaccgtg caaaaggaga gcctgtaagc gggcactctt ccgtggtctg gtggataaat
     5041 tcgcaagggt atcatggcgg acgaccgggg ttcgaacccc ggatccggcc gtccgccgtg
     5101 atccatgcgg ttaccgcccg cgtgtcgaac ccaggtgtgc gacgtcagac aacgggggag
     5161 cgctcctttt ggcttccttc caggcgcggc ggctgctgcg ctagcttttt tggccactgg
     5221 ccgcgcgcgg cgtaagcggt taggctgcca tctctatcgg tatcgtagca taaccccttg
     5281 gggcctctaa acgggtcttg aggggttttt tgtgcccctc gggccggatt gctatctacc
     5341 ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg
     5401 ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta
     5461 ccagaaattt atccttaagg tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa
     5521 gacattcctt taatggtctt ttctggacac cactaggggt cagaagtagt tcatcaaact
     5581 ttcttccctc cctaatctca ttggttacct tgggctatcg aaacttaatt aaccagtcaa
     5641 gtcagctact tggcgagatc gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt
     5701 caagttcgat ctggtccttg ctattgcacc cgttctccga ttacgagttt catttaaatc
     5761 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5821 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5881 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5941 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6001 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6061 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6121 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6181 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6241 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6301 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6361 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6421 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6481 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6541 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6601 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcatttaa atttccgaac
     6661 tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc cgatccatct tgcaggctac
     6721 ctctcgaacg aactatcgca agtctcttgg ccggccttgc gccttggcta ttgcttggca
     6781 gcgcctatcg ccaggtatta ctccaatccc gaatatccga gatcgggatc acccgagaga
     6841 agttcaacct acatcctcaa tcccgatcta tccgagatcc gaggaatatc gaaatcgggg
     6901 cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc
     6961 ttggcagtca gccagtcgga atccagcttg ggacccagga agtccaatcg tcagatattg
     7021 tactcaagcc tggtcacggc agcgtaccga tctgtttaaa cctagatatt gatagtctga
     7081 tcggtcaacg tataatcgag tcctagcttt tgcaaacatc tatcaagaga caggatcagc
     7141 aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     7201 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     7261 tcagttgggt gcgcgagtgg gttacatcga actggatctc aacagcggta agatccttga
     7321 gagttttcgc cccgaagaac gctttccaat gatgagcact tttaaagttc tgctatgtgg
     7381 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     7441 tcagaatgac ttggttgagt attcaccagt cacagaaaag catcttacgg atggcatgac
     7501 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     7561 tctgacaacg attggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     7621 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     7681 tgacaccacg atgcctgtag caatggcaac aaccttgcgt aaactattaa ctggcgaact
     7741 acttactcta gcttcccggc aacagttgat agactggatg gaggcggata aagttgcagg
     7801 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     7861 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     7921 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     7981 tgagataggt gcctcactga ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa
     8041 aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg
     8101 gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct
     8161 taagatcccg aatcgtttaa actcgactct ggctctatcg aatctccgtc gtttcgagct
     8221 tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc gaa
//
