LOCUS       sequence-455021-        5534 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ef1a promoter driving expression of adenoviral L4-22k gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5534)
  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 5534)
  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: JD182
FEATURES             Location/Qualifiers
     source          1..5534
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(46..92)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    110..158
                     /note="synthetic polyadenylation signal"
     promoter        195..1373
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          426..1364
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1321..1341
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1579..1588
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2219..2340
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2230..2249)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2284..2303
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2455..2501
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2531..2667
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2712..2783
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3009..3597)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3089..3108)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4343..4411,4412..5203)
                     /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(4561..4580)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5216..5352
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    join(5504..5534,1..41)
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
ORIGIN
        1 cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa
       61 gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa
      121 ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta
      181 gccagatctc gtgaggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc
      241 ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg
      301 taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac
      361 cgtatataag tgcactagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa
      421 cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt tatggccctt
      481 gcgtgccttg aattacttcc acctggctgc agtacgtgat tcttgatccc gagcttcggg
      541 ttggaagtgg gtgggagagt tcgtggcctt gcgcttaagg agccccttcg cctcgtgctt
      601 gagttgtggc ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg
      661 cctgtctcgc tgctttcgat aagtctctag ccatttaaaa tttttgatga cctgctgcga
      721 cgcttttttt ctggcaagat agtcttgtaa atgcgggcca agatcagcac actggtattt
      781 cggtttttgg ggccgcgggc ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga
      841 ggcggggcct gcgagcgcgg ccaccgagaa tcggacgggg gtagtctcaa gctgcccggc
      901 ctgctctggt gcctggcctc gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg
      961 cccggtcggc accagttgcg tgagcggaaa gatggccgct tcccggccct gctgcaggga
     1021 gcacaaaatg gaggacgcgg cgctcgggag agcgggcggg tgagtcaccc acacaaagga
     1081 aaagggcctt tccgtcctca gccgtcgctt catgtgactc cacggagtac cgggcgccgt
     1141 ccaggcacct cgattagttc tccagctttt ggagtacgtc gtctttaggt tggggggagg
     1201 ggttttatgc gatggagttt ccccacactg agtgggtgga gactgaagtt aggccagctt
     1261 ggcacttgat gtaattctcc ttggaatttg ccctttttga gtttggatct tggttcattc
     1321 tcaagcctca gacagtggtt caaagttttt ttcttccatt tcaggtgtcg tgaaaactac
     1381 ccctaaaagc caaaagatct ttgtcgatcc taccatccac tcgacacacc cgccagcggc
     1441 cgctgccaag cttccgagct ctcgaattca tcttggctga ggaatcttct aacaatttag
     1501 agcttaaaaa cgcccacgag gcggagaacg aaatatccag agagacgtta gaaacgttca
     1561 aaaacgttca ccggtagggc caccatggca cccaaaaaga agctgcagct gccgccgcca
     1621 cccacggacg aggaggaata ctgggacagt caggcagagg aggttttgga cgaggaggag
     1681 gaggacatga tggaagactg ggagagccta gacgaggaag cttccgaggt cgaagaggtg
     1741 tcagacgaaa caccgtcacc ctcggtcgca ttcccctcgc cggcgcccca gaaatcggca
     1801 accggttcca gcatggctac aacctccgct cctcaggcgc cgccggcact gcccgttcgc
     1861 cgacccaacc gtagatggga caccactgga accagggccg gtaagtccaa gcagccgccg
     1921 ccgttagccc aagagcaaca acagcgccaa ggctaccgct catggcgcgg gcacaagaac
     1981 gccatagttg cttgcttgca agactgtggg ggcaacatct ccttcgcccg ccgctttctt
     2041 ctctaccatc acggcgtggc cttcccccgt aacatcctgc attactaccg tcatctctac
     2101 agcccatact gcaccggcgg cagcggcagc aacagcagcg gccacacaga agcaaaggcg
     2161 accggatagg ctagcttgac tgactgagat acagcgtacc ttcagctcac agacatgata
     2221 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     2281 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     2341 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     2401 taaagcaagt aaaacctcta caaatgtggt attggcccat ctctatcggt atcgtagcat
     2461 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg ggccggattg
     2521 ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc
     2581 ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg
     2641 tcctccttac cagaaattta tccttaaggt cgtcagctat cctgcaggcg atctctcgat
     2701 ttcgatcaag acattccttt aatggtcttt tctggacacc actaggggtc agaagtagtt
     2761 catcaaactt tcttccctcc ctaatctcat tggttacctt gggctatcga aacttaatta
     2821 accagtcaag tcagctactt ggcgagatcg acttgtctgg gtttcgacta cgctcagaat
     2881 tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc gttctccgat tacgagtttc
     2941 atttaaatca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3001 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3061 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3121 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3181 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3241 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3301 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3361 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3421 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3481 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3541 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3601 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3661 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3721 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3781 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcatttaaa
     3841 tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt
     3901 gcaggctacc tctcgaacga actatcgcaa gtctcttggc cggccttgcg ccttggctat
     3961 tgcttggcag cgcctatcgc caggtattac tccaatcccg aatatccgag atcgggatca
     4021 cccgagagaa gttcaaccta catcctcaat cccgatctat ccgagatccg aggaatatcg
     4081 aaatcggggc gcgcctggtg taccgagaac gatcctctca gtgcgagtct cgacgatcca
     4141 tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt
     4201 cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac ctagatattg
     4261 atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct atcaagagac
     4321 aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4381 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4441 tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca acagcggtaa
     4501 gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt ttaaagttct
     4561 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4621 acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc atcttacgga
     4681 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4741 caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt tgcacaacat
     4801 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4861 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta aactattaac
     4921 tggcgaacta cttactctag cttcccggca acagttgata gactggatgg aggcggataa
     4981 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5041 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5101 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5161 acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc taggtgcatt
     5221 ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat
     5281 tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa
     5341 atttatcctt aagatcccga atcgtttaaa ctcgactctg gctctatcga atctccgtcg
     5401 tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg aatctcgtca
     5461 gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg gaccattagc
     5521 tccacaggta tctt
//
