LOCUS       sequence-455007-        5628 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ef1a promoter driving expression of adenoviral L4-33k gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5628)
  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 5628)
  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: JD183
FEATURES             Location/Qualifiers
     source          1..5628
                     /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    2313..2434
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2324..2343)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2378..2397
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2549..2595
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2625..2761
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2806..2877
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3103..3691)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3183..3202)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4437..4505,4506..5297)
                     /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(4655..4674)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5310..5446
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    join(5598..5628,1..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 cccatactgc accggcggca
     1921 gcggcagcaa cagcagcggc cacacagaag caaaggcgac cggatagcaa gactctgaca
     1981 aagcccaaga aatccacagc ggcggcagca gcaggaggag gagcgctgcg tctggcgccc
     2041 aacgaacccg tatcgacccg cgagcttaga aacaggattt ttcccactct gtatgctata
     2101 tttcaacaga gcaggggcca agaacaagag ctgaaaataa aaaacaggtc tctgcgatcc
     2161 ctcacccgca gctgcctgta tcacaaaagc gaagatcagc ttcggcgcac gctggaagac
     2221 gcggaggctc tcttcagtaa atactgcgcg ctgactctta aggactagct tgactgactg
     2281 agatacagcg taccttcagc tcacagacat gataagatac attgatgagt ttggacaaac
     2341 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2401 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2461 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     2521 tggtattggc ccatctctat cggtatcgta gcataacccc ttggggcctc taaacgggtc
     2581 ttgaggggtt ttttgtgccc ctcgggccgg attgctatct accggcattg gcgcagaaaa
     2641 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     2701 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     2761 aggtcgtcag ctatcctgca ggcgatctct cgatttcgat caagacattc ctttaatggt
     2821 cttttctgga caccactagg ggtcagaagt agttcatcaa actttcttcc ctccctaatc
     2881 tcattggtta ccttgggcta tcgaaactta attaaccagt caagtcagct acttggcgag
     2941 atcgacttgt ctgggtttcg actacgctca gaattgcgtc agtcaagttc gatctggtcc
     3001 ttgctattgc acccgttctc cgattacgag tttcatttaa atcatgtgag caaaaggcca
     3061 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3121 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3181 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3241 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3301 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3361 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3421 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3481 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3541 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3601 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3661 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3721 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3781 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3841 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3901 ctgtctattt cgttcatcca tagttgcatt taaatttccg aactctccaa ggccctcgtc
     3961 ggaaaatctt caaacctttc gtccgatcca tcttgcaggc tacctctcga acgaactatc
     4021 gcaagtctct tggccggcct tgcgccttgg ctattgcttg gcagcgccta tcgccaggta
     4081 ttactccaat cccgaatatc cgagatcggg atcacccgag agaagttcaa cctacatcct
     4141 caatcccgat ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct ggtgtaccga
     4201 gaacgatcct ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc
     4261 ggaatccagc ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac
     4321 ggcagcgtac cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc
     4381 gagtcctagc ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga
     4441 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4501 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag
     4561 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4621 aacgctttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     4681 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4741 agtattcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     4801 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgattggag
     4861 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     4921 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     4981 tagcaatggc aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc
     5041 ggcaacagtt gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     5101 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     5161 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     5221 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     5281 tgattaagca ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga
     5341 cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac
     5401 ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaagatc ccgaatcgtt
     5461 taaactcgac tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg
     5521 cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca
     5581 gcgatcgcgg ctcccgacat cttggaccat tagctccaca ggtatctt
//
