LOCUS       sequence-455009-        8564 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Adenoviral helper plasmid containing E2A, VA RNA I and L4-22k/33k 
            genes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8564)
  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 8564)
  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: JD171
FEATURES             Location/Qualifiers
     source          1..8564
                     /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    2602..2723
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2613..2632)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2667..2686
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    2732..2956
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(4712..4915)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4741..4761)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4916..5219
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     terminator      5567..5613
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      5643..5779
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5824..5895
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(6121..6709)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6201..6220)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(7455..7523,7524..8315)
                     /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(7673..7692)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      8328..8464
                     /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 gcaagactct gacaaagccc aagaaatcca
     2281 cagcggcggc agcagcagga ggaggagcgc tgcgtctggc gcccaacgaa cccgtatcga
     2341 cccgcgagct tagaaacagg atttttccca ctctgtatgc tatatttcaa cagagcaggg
     2401 gccaagaaca agagctgaaa ataaaaaaca ggtctctgcg atccctcacc cgcagctgcc
     2461 tgtatcacaa aagcgaagat cagcttcggc gcacgctgga agacgcggag gctctcttca
     2521 gtaaatactg cgcgctgact cttaaggact aggctagctt gactgactga gatacagcgt
     2581 accttcagct cacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     2641 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     2701 ttataagctg caataaacaa gttaggaatg gccatagagc ccaccgcatc cccagcatgc
     2761 ctgctattgt gttcccaatc ctcccccttg ctgtcctgcc ccaccccacc ccccagaata
     2821 gaatgacacc tactcagaca atgcgatgca atttcctcat tttattagga aaggacagtg
     2881 ggagtggcac cttccagggt caaggaaggc acgggggagg ggcaaacaac agatggctgg
     2941 caactagaag gcacagtaaa atagataaat ggcatcttgc cgtctgcgcc gtttaaaaat
     3001 caaaggggtt ctgccgcgca tcgctatgcg ccactggcag ggacacgttg cgatactggt
     3061 gtttagtgct ccacttaaac tcaggcacaa ccatccgcgg cagctcggtg aagttttcac
     3121 tccacaggct gcgcaccatc accaacgcgt ttagcaggtc gggcgccgat atcttgaagt
     3181 cgcagttggg gcctccgccc tgcgcgcgcg agttgcgata cacagggttg cagcactgga
     3241 acactatcag cgccgggtgg tgcacgctgg ccagcacgct cttgtcggag atcagatccg
     3301 cgtccaggtc ctccgcgttg ctcagggcga acggagtcaa ctttggtagc tgccttccca
     3361 aaaagggtgc atgcccaggc tttgagttgc actcgcaccg tagtggcatc agaaggtgac
     3421 cgtgcccggt ctgggcgtta ggatacagcg cctgcatgaa agccttgatc tgcttaaaag
     3481 ccacctgagc ctttgcgcct tcagagaaga acatgccgca agacttgccg gaaaactgat
     3541 tggccggaca ggccgcgtca tgcacgcagc accttgcgtc ggtgttggag atctgcacca
     3601 catttcggcc ccaccggttc ttcacgatct tggccttgct agactgctcc ttcagcgcgc
     3661 gctgcccgtt ttcgctcgtc acatccattt caatcacgtg ctccttattt atcataatgc
     3721 tcccgtgtag acacttaagc tcgccttcga tctcagcgca gcggtgcagc cacaacgcgc
     3781 agcccgtggg ctcgtggtgc ttgtaggtta cctctgcaaa cgactgcagg tacgcctgca
     3841 ggaatcgccc catcatcgtc acaaaggtct tgttgctggt gaaggtcagc tgcaacccgc
     3901 ggtgctcctc gtttagccag gtcttgcata cggccgccag agcttccact tggtcaggca
     3961 gtagcttgaa gtttgccttt agatcgttat ccacgtggta cttgtccatc aacgcgcgcg
     4021 cagcctccat gcccttctcc cacgcagaca cgatcggcag gctcagcggg tttatcaccg
     4081 tgctttcact ttccgcttca ctggactctt ccttttcctc ttgcgtccgc ataccccgcg
     4141 ccactgggtc gtcctcattc agccgccgca ccgtgcgctt acctcccttg ccgtgcttga
     4201 ttagcaccgg tgggttgctg aaacccacca tttgtagcgc cacatcttct ctttcttcct
     4261 cgctgtccac gatcacctct ggggatggcg ggcgctcggg cttgggagag gggcgcttct
     4321 ttttcttttt ggacgcaatg gccaaatccg ccgtcgaggt cgatggccgc gggctgggtg
     4381 tgcgcggcac cagcgcatct tgtgacgagt cctcttcgtc ctcggactcg agccgccgcc
     4441 tcagccgctt ttttgggggc gcgcggggag gcggcggcga cggcgacggg gacgacacgt
     4501 cctccatggt tggtggacgt cgcgccgcac cgcgtccgcg ctcgggggtg gtttcgcgct
     4561 gctcctcttc ccgactggcc atgctggcgg taccttcctt ctcctgatat cgcctcgctc
     4621 gacgaagtgc caaaaatctt tgagggtctt ggacgcgacg agaaacgcgc ggcaaacgct
     4681 ccgcagctgc ctgtatcaca acctaggtct gagctctgct tatatagacc tcccaccgta
     4741 cacgcctacc gcccatttgc gtcaatgggg cggagttgtt acgacatttt ggaaagtccc
     4801 gttgattttg gtgccaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc
     4861 cccgtgagtc aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg
     4921 taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt
     4981 actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg
     5041 gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt
     5101 gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca
     5161 atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc
     5221 tccgtaccag ctccatggtc gggacgctct ggccggtgag gcgtgcgcag tcgttgacgc
     5281 tctagaccgt gcaaaaggag agcctgtaag cgggcactct tccgtggtct ggtggataaa
     5341 ttcgcaaggg tatcatggcg gacgaccggg gttcgaaccc cggatccggc cgtccgccgt
     5401 gatccatgcg gttaccgccc gcgtgtcgaa cccaggtgtg cgacgtcaga caacggggga
     5461 gcgctccttt tggcttcctt ccaggcgcgg cggctgctgc gctagctttt ttggccactg
     5521 gccgcgcgcg gcgtaagcgg ttaggctgcc atctctatcg gtatcgtagc ataacccctt
     5581 ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat tgctatctac
     5641 cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     5701 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     5761 accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg atttcgatca
     5821 agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag ttcatcaaac
     5881 tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat taaccagtca
     5941 agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga attgcgtcag
     6001 tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt tcatttaaat
     6061 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6121 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6181 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6241 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6301 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6361 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6421 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6481 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6541 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6601 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6661 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6721 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6781 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6841 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6901 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta aatttccgaa
     6961 ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc ttgcaggcta
     7021 cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct attgcttggc
     7081 agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat cacccgagag
     7141 aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat cgaaatcggg
     7201 gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt ctcgacgatc catatcgttg
     7261 cttggcagtc agccagtcgg aatccagctt gggacccagg aagtccaatc gtcagatatt
     7321 gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa acctagatat tgatagtctg
     7381 atcggtcaac gtataatcga gtcctagctt ttgcaaacat ctatcaagag acaggatcag
     7441 caggaggctt tcgcatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     7501 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     7561 atcagttggg tgcgcgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     7621 agagttttcg ccccgaagaa cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg
     7681 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     7741 ctcagaatga cttggttgag tattcaccag tcacagaaaa gcatcttacg gatggcatga
     7801 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     7861 ttctgacaac gattggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     7921 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     7981 gtgacaccac gatgcctgta gcaatggcaa caaccttgcg taaactatta actggcgaac
     8041 tacttactct agcttcccgg caacagttga tagactggat ggaggcggat aaagttgcag
     8101 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     8161 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     8221 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     8281 ctgagatagg tgcctcactg attaagcatt ggtaaccgat tctaggtgca ttggcgcaga
     8341 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
     8401 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
     8461 ttaagatccc gaatcgttta aactcgactc tggctctatc gaatctccgt cgtttcgagc
     8521 ttacgcgaac agccgtggcg ctcatttgct cgtcgggcat cgaa
//
