LOCUS       sequence-417470-        6371 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses DNAJC11 fused with EGFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6371)
  AUTHORS   Li J, Gui Q, Liang FX, Sall J, Zhang Q, Duan Y, Dhabaria A, Askenazi
            M, Ueberheide B, Stapleford KA, Pagano M
  TITLE     The REEP5/TRAM1 complex binds SARS-CoV-2 NSP3 and promotes virus 
            replication.
  JOURNAL   J Virol. 2023 Sep 28:e0050723. doi: 10.1128/jvi.00507-23.
  PUBMED    37768083
REFERENCE   2  (bases 1 to 6371)
  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: "J Virol. 
            2023 Sep 28:e0050723. doi: 10.1128/jvi.00507-23."
COMMENT     Sequence Label: DNAJC11-EGFP
FEATURES             Location/Qualifiers
     source          1..6371
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        52..355
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        356..559
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     510..530
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2302..2311
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2308..2310,2311..2313,2314..3027)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2353..2374)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2614..2633)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2961..2982
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3150..3271
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3187..3206)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3241..3260
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3278..3733)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3415..3436)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3627..3646
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3760..3864
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3866..4223
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3866..4057
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3889..3909)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4074..4209
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4258..5052
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4258..5052
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4312..4331)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4922..4941
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5240..5259)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5284..5331
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5660..6248
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6149..6168
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
       61 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      121 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      181 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      241 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      301 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      361 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      421 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      481 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      541 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggactca
      601 gatctcgaga tggcgacggc cttgagcgag gaggagctgg acaatgaaga ctattactcg
      661 ttgctgaacg tgcgcaggga ggcctcttct gaagagctga aagctgccta ccggaggctc
      721 tgtatgctct accatccaga caagcacaga gacccagagc tcaagtcaca ggcggaacga
      781 ctgtttaacc ttgttcacca ggcttatgaa gtgcttagtg acccccaaac cagggccatc
      841 tatgatatat atgggaagag aggactggaa atggaaggat gggaggttgt ggaaaggagg
      901 agaacccctg ctgaaattcg agaggagttt gagcggctgc agagagagag agaagagagg
      961 agattgcagc agcgaaccaa tcccaaggga acgatcagcg ttggagtaga tgccaccgac
     1021 ctttttgatc gctatgatga ggagtatgaa gatgtgtccg gcagtagctt tccgcagatt
     1081 gaaattaata aaatgcacat atcccagtcc attgaggcac ccttgacagc gacagacaca
     1141 gccatcctct ctggaagcct ctcaacccag aatggaaatg gaggaggttc cattaacttt
     1201 gcgctcagac gagtaacttc ggcaaaggga tggggagagt tggaatttgg agctggagac
     1261 ctacaggggc ctttgttcgg tctcaagctg ttccgtaatc tcacaccaag atgctttgtg
     1321 acaacaaact gtgctctgca gttttcatcc cgtggaatcc gacccggcct gaccactgtc
     1381 ctagctcgga acctagacaa gaacaccgtg ggctacctgc agtggcgatg gggtatccag
     1441 tcagccatga acactagcat cgtccgagac actaaaagca gccacttcac tgtggccctg
     1501 cagctgggaa tccctcactc ctttgcactg atcagctatc agcacaaatt ccaagatgac
     1561 gatcagactc gtgtgaaagg atccctcaaa gcaggcttct ttgggacggt ggtggagtac
     1621 ggagctgaga ggaagatctc caggcacagc gttttgggtg cagctgtcag cgttggagtt
     1681 ccacagggcg tttctctcaa agtcaagctc aacagggcca gtcagacata cttcttccct
     1741 attcacttga cggaccagct tctgcccagc gccatgttct atgccaccgt ggggcctcta
     1801 gtggtctact ttgccatgca ccgtctgatc atcaaaccat acctcagggc tcagaaagag
     1861 aaggaattgg agaagcagag ggaaagcgcc gccaccgatg tgctgcagaa gaagcaagag
     1921 gcggagtccg ctgtccggct gatgcaggaa tctgtccgaa ggataattga ggcagaagag
     1981 tccagaatgg gcctcatcat cgtcaatgcc tggtacggga agtttgtcaa tgacaagagc
     2041 aggaagagcg agaaggtgaa ggtgattgac gtgactgtgc ccctgcagtg cctggtgaag
     2101 gactcgaagc tcatcctcac ggaggcctcc aaggctgggc tgcctggctt ttatgacccg
     2161 tgtgtggggg aagagaagaa cctgaaagtg ctctatcagt tccggggcgt cctgcatcag
     2221 gtgatggtgc tggacagtga ggccctccgg ataccaaagc agtcccacag gatcgataca
     2281 gatggagcgg atccaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
     2341 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     2401 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     2461 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     2521 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     2581 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     2641 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     2701 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     2761 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     2821 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     2881 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccaagct gagcaaagac
     2941 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     3001 ctcggcatgg acgagctgta caagtaaagc ggccgcgact ctagatcata atcagccata
     3061 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     3121 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     3181 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     3241 gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg
     3301 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     3361 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     3421 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     3481 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     3541 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     3601 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     3661 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     3721 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3781 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3841 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     3901 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3961 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4021 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4081 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4141 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     4201 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
     4261 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4321 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4381 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     4441 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4501 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4561 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4621 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4681 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4741 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     4801 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4861 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4921 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4981 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5041 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     5101 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     5161 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     5221 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     5281 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
     5341 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     5401 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     5461 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
     5521 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5581 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5641 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5701 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5761 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5821 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5881 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5941 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6001 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6061 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6121 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6181 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6241 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6301 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc
     6361 attagttatt a
//
