LOCUS       sequence-456052-        7374 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ef1a promoter driving expression of adenoviral 100k gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7374)
  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 7374)
  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: JD135
FEATURES             Location/Qualifiers
     source          1..7374
                     /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    4059..4180
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4070..4089)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4124..4143
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      4295..4341
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4371..4507
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4552..4623
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4849..5437)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4929..4948)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(6183..6251,6252..7043)
                     /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(6401..6420)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7056..7192
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    join(7344..7374,1..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 caccatggag tcagtcgaga agaaggacag cctaaccgcc
     1621 ccctctgagt tcgccaccac cgcctccacc gatgccgcca acgcgcctac caccttcccc
     1681 gtcgaggcac ccccgcttga ggaggaggaa gtgattatcg agcaggaccc aggttttgta
     1741 agcgaagacg acgaggaccg ctcagtacca acagaggata aaaagcaaga ccaggacaac
     1801 gcagaggcaa acgaggaaca agtcgggcgg ggggacgaaa ggcatggcga ctacctagat
     1861 gtgggagacg acgtgctgtt gaagcatctg cagcgccagt gcgccattat ctgcgacgcg
     1921 ttgcaagagc gcagcgatgt gcccctcgcc atagcggatg tcagccttgc ctacgaacgc
     1981 cacctattct caccgcgcgt accccccaaa cgccaagaaa acggcacatg cgagcccaac
     2041 ccgcgcctca acttctaccc cgtatttgcc gtgccagagg tgcttgccac ctatcacatc
     2101 tttttccaaa actgcaagat acccctatcc tgccgtgcca accgcagccg agcggacaag
     2161 cagctggcct tgcggcaggg cgctgtcata cctgatatcg cctcgctcaa cgaagtgcca
     2221 aaaatctttg agggtcttgg acgcgacgag aagcgcgcgg caaacgctct gcaacaggaa
     2281 aacagcgaaa atgaaagtca ctctggagtg ttggtggaac tcgagggtga caacgcgcgc
     2341 ctagccgtac taaaacgcag catcgaggtc acccactttg cctacccggc acttaaccta
     2401 ccccccaagg tcatgagcac agtcatgagt gagctgatcg tgcgccgtgc gcagcccctg
     2461 gagagggatg caaatttgca agaacaaaca gaggagggcc tacccgcagt tggcgacgag
     2521 cagctagcgc gctggcttca aacgcgcgag cctgccgact tggaggagcg acgcaaacta
     2581 atgatggccg cagtgctcgt taccgtggag cttgagtgca tgcagcggtt ctttgctgac
     2641 ccggagatgc agcgcaagct agaggaaaca ttgcactaca cctttcgaca gggctacgta
     2701 cgccaggcct gcaagatctc caacgtggag ctctgcaacc tggtctccta ccttggaatt
     2761 ttgcacgaaa accgccttgg gcaaaacgtg cttcattcca cgctcaaggg cgaggcgcgc
     2821 cgcgactacg tccgcgactg cgtttactta tttctatgct acacctggca gacggccatg
     2881 ggcgtttggc agcagtgctt ggaggagtgc aacctcaagg agctgcagaa actgctaaag
     2941 caaaacttga aggacctatg gacggccttc aacgagcgct ccgtggccgc gcacctggcg
     3001 gacatcattt tccccgaacg cctgcttaaa accctgcaac agggtctgcc agacttcacc
     3061 agtcaaagca tgttgcagaa ctttaggaac tttatcctag agcgctcagg aatcttgccc
     3121 gccacctgct gtgcacttcc tagcgacttt gtgcccatta agtaccgcga atgccctccg
     3181 ccgctttggg gccactgcta ccttctgcag ctagccaact accttgccta ccactctgac
     3241 ataatggaag acgtgagcgg tgacggtcta ctggagtgtc actgtcgctg caacctatgc
     3301 accccgcacc gctccctggt ttgcaattcg cagctgctta acgaaagtca aattatcggt
     3361 acctttgagc tgcagggtcc ctcgcctgac gaaaagtccg cggctccggg gttgaaactc
     3421 actccggggc tgtggacgtc ggcttacctt cgcaaatttg tacctgagga ctaccacgcc
     3481 cacgagatta ggttctacga agaccaatcc cgcccgccta atgcggagct taccgcctgc
     3541 gtcattaccc agggccacat tcttggccaa ttgcaagcca tcaacaaagc ccgccaagag
     3601 tttctgctac gaaagggacg gggggtttac ttggaccccc agtccggcga ggagctcaac
     3661 ccaatccccc cgccgccgca gccctatcag cagcagccgc gggcccttgc ttcccaggat
     3721 ggcacccaaa aagaagctgc agctgccgcc gccacccacg gacgaggagg aatactggga
     3781 cagtcaggca gaggaggttt tggacgagga ggaggaggac atgatggaag actgggagag
     3841 cctagacgag gaagcttccg aggtcgaaga ggtgtcagac gaaacaccgt caccctcggt
     3901 cgcattcccc tcgccggcgc cccagaaatc ggcaaccggt tccagcatgg ctacaacctc
     3961 cgctcctcag gcgccgccgg cactgcccgt tcgccgaccc aaccgtagcc taggcttgac
     4021 tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg
     4081 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     4141 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     4201 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     4261 caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa
     4321 cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc
     4381 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     4441 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     4501 tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt
     4561 aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc
     4621 ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt
     4681 ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc
     4741 tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa
     4801 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4861 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4921 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4981 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5041 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5101 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5161 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5221 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5281 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5341 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5401 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5461 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5521 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5581 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5641 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc
     5701 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
     5761 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
     5821 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
     5881 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggtg
     5941 taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag
     6001 ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct
     6061 ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt
     6121 ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc
     6181 gcatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     6241 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     6301 cgcgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     6361 ccgaagaacg ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     6421 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     6481 tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     6541 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     6601 ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     6661 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     6721 tgcctgtagc aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag
     6781 cttcccggca acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc
     6841 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     6901 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     6961 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     7021 cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg
     7081 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     7141 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcccga
     7201 atcgtttaaa ctcgactctg gctctatcga atctccgtcg tttcgagctt acgcgaacag
     7261 ccgtggcgct catttgctcg tcgggcatcg aatctcgtca gctatcgtca gcttaccttt
     7321 ttggcagcga tcgcggctcc cgacatcttg gaccattagc tccacaggta tctt
//
