LOCUS       sequence-455017-        5835 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ef1a promoter driving expression of adenoviral L4-22k/33k genes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5835)
  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 5835)
  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: JD141
FEATURES             Location/Qualifiers
     source          1..5835
                     /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    2520..2641
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2531..2550)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2585..2604
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2756..2802
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2832..2968
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3013..3084
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3310..3898)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3390..3409)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4644..4712,4713..5504)
                     /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(4862..4881)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5517..5653
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    join(5805..5835,1..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 accggatagc aagactctga caaagcccaa gaaatccaca gcggcggcag cagcaggagg
     2221 aggagcgctg cgtctggcgc ccaacgaacc cgtatcgacc cgcgagctta gaaacaggat
     2281 ttttcccact ctgtatgcta tatttcaaca gagcaggggc caagaacaag agctgaaaat
     2341 aaaaaacagg tctctgcgat ccctcacccg cagctgcctg tatcacaaaa gcgaagatca
     2401 gcttcggcgc acgctggaag acgcggaggc tctcttcagt aaatactgcg cgctgactct
     2461 taaggactag gctagcttga ctgactgaga tacagcgtac cttcagctca cagacatgat
     2521 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     2581 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     2641 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     2701 ttaaagcaag taaaacctct acaaatgtgg tattggccca tctctatcgg tatcgtagca
     2761 taaccccttg gggcctctaa acgggtcttg aggggttttt tgtgcccctc gggccggatt
     2821 gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
     2881 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
     2941 gtcctcctta ccagaaattt atccttaagg tcgtcagcta tcctgcaggc gatctctcga
     3001 tttcgatcaa gacattcctt taatggtctt ttctggacac cactaggggt cagaagtagt
     3061 tcatcaaact ttcttccctc cctaatctca ttggttacct tgggctatcg aaacttaatt
     3121 aaccagtcaa gtcagctact tggcgagatc gacttgtctg ggtttcgact acgctcagaa
     3181 ttgcgtcagt caagttcgat ctggtccttg ctattgcacc cgttctccga ttacgagttt
     3241 catttaaatc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3301 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3361 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3421 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3481 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3541 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3601 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3661 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3721 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3781 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3841 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3901 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3961 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4021 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4081 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcatttaa
     4141 atttccgaac tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc cgatccatct
     4201 tgcaggctac ctctcgaacg aactatcgca agtctcttgg ccggccttgc gccttggcta
     4261 ttgcttggca gcgcctatcg ccaggtatta ctccaatccc gaatatccga gatcgggatc
     4321 acccgagaga agttcaacct acatcctcaa tcccgatcta tccgagatcc gaggaatatc
     4381 gaaatcgggg cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc tcgacgatcc
     4441 atatcgttgc ttggcagtca gccagtcgga atccagcttg ggacccagga agtccaatcg
     4501 tcagatattg tactcaagcc tggtcacggc agcgtaccga tctgtttaaa cctagatatt
     4561 gatagtctga tcggtcaacg tataatcgag tcctagcttt tgcaaacatc tatcaagaga
     4621 caggatcagc aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc cttattccct
     4681 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4741 atgctgaaga tcagttgggt gcgcgagtgg gttacatcga actggatctc aacagcggta
     4801 agatccttga gagttttcgc cccgaagaac gctttccaat gatgagcact tttaaagttc
     4861 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     4921 tacactattc tcagaatgac ttggttgagt attcaccagt cacagaaaag catcttacgg
     4981 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5041 ccaacttact tctgacaacg attggaggac cgaaggagct aaccgctttt ttgcacaaca
     5101 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5161 acgacgagcg tgacaccacg atgcctgtag caatggcaac aaccttgcgt aaactattaa
     5221 ctggcgaact acttactcta gcttcccggc aacagttgat agactggatg gaggcggata
     5281 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5341 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5401 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5461 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaaccgatt ctaggtgcat
     5521 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
     5581 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
     5641 aatttatcct taagatcccg aatcgtttaa actcgactct ggctctatcg aatctccgtc
     5701 gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc gaatctcgtc
     5761 agctatcgtc agcttacctt tttggcagcg atcgcggctc ccgacatctt ggaccattag
     5821 ctccacaggt atctt
//
