LOCUS       sequence-417467-        7229 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses DNAJC11 fused with 2Strep in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7229)
  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 7229)
  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: DNAJC11-2Strep
FEATURES             Location/Qualifiers
     source          1..7229
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(32..51)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        223..602
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        603..806
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     757..777
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     851..870
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        851..869
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2587..2596
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2591..2600
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2608..2631
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2668..2691
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2716..2739
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     promoter        complement(2770..2788)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2771..2788)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2808..2825)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2814..3038
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3084..3512
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3171..3190)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3381..3402
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3521..3541)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3526..3855
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3706..3841
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3768..3787
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3922..4716
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3976..3995)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4586..4605
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4890..5011
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4927..4946)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4981..5000
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5060..5076)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5060..5076)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5073..5095)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5084..5100
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5108..5114,5115..5132,5133..5138))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5153..5174
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5291..5308)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5462..6050)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5542..5561)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6221..7012,7013..7081))
                     /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     6844..6863
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7082..7186)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
       61 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
      121 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
      181 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta
      241 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      301 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      361 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      421 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      481 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      541 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      601 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      661 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      721 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      781 ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact gcttactggc
      841 ttatcgaaat taatacgact cactataggg agacccaagc ttggtaccga gctcggatcc
      901 atggcgacgg ccttgagcga ggaggagctg gacaatgaag actattactc gttgctgaac
      961 gtgcgcaggg aggcctcttc tgaagagctg aaagctgcct accggaggct ctgtatgctc
     1021 taccatccag acaagcacag agacccagag ctcaagtcac aggcggaacg actgtttaac
     1081 cttgttcacc aggcttatga agtgcttagt gacccccaaa ccagggccat ctatgatata
     1141 tatgggaaga gaggactgga aatggaagga tgggaggttg tggaaaggag gagaacccct
     1201 gctgaaattc gagaggagtt tgagcggctg cagagagaga gagaagagag gagattgcag
     1261 cagcgaacca atcccaaggg aacgatcagc gttggagtag atgccaccga cctttttgat
     1321 cgctatgatg aggagtatga agatgtgtcc ggcagtagct ttccgcagat tgaaattaat
     1381 aaaatgcaca tatcccagtc cattgaggca cccttgacag cgacagacac agccatcctc
     1441 tctggaagcc tctcaaccca gaatggaaat ggaggaggtt ccattaactt tgcgctcaga
     1501 cgagtaactt cggcaaaggg atggggagag ttggaatttg gagctggaga cctacagggg
     1561 cctttgttcg gtctcaagct gttccgtaat ctcacaccaa gatgctttgt gacaacaaac
     1621 tgtgctctgc agttttcatc ccgtggaatc cgacccggcc tgaccactgt cctagctcgg
     1681 aacctagaca agaacaccgt gggctacctg cagtggcgat ggggtatcca gtcagccatg
     1741 aacactagca tcgtccgaga cactaaaagc agccacttca ctgtggccct gcagctggga
     1801 atccctcact cctttgcact gatcagctat cagcacaaat tccaagatga cgatcagact
     1861 cgtgtgaaag gatccctcaa agcaggcttc tttgggacgg tggtggagta cggagctgag
     1921 aggaagatct ccaggcacag cgttttgggt gcagctgtca gcgttggagt tccacagggc
     1981 gtttctctca aagtcaagct caacagggcc agtcagacat acttcttccc tattcacttg
     2041 acggaccagc ttctgcccag cgccatgttc tatgccaccg tggggcctct agtggtctac
     2101 tttgccatgc accgtctgat catcaaacca tacctcaggg ctcagaaaga gaaggaattg
     2161 gagaagcaga gggaaagcgc cgccaccgat gtgctgcaga agaagcaaga ggcggagtcc
     2221 gctgtccggc tgatgcagga atctgtccga aggataattg aggcagaaga gtccagaatg
     2281 ggcctcatca tcgtcaatgc ctggtacggg aagtttgtca atgacaagag caggaagagc
     2341 gagaaggtga aggtgattga cgtgactgtg cccctgcagt gcctggtgaa ggactcgaag
     2401 ctcatcctca cggaggcctc caaggctggg ctgcctggct tttatgaccc gtgtgtgggg
     2461 gaagagaaga acctgaaagt gctctatcag ttccggggcg tcctgcatca ggtgatggtg
     2521 ctggacagtg aggccctccg gataccaaag cagtcccaca ggatcgatac agatggagaa
     2581 ttcgccgcca ccatggcggc cgccagctgg agccaccctc agttcgagaa gggaggagga
     2641 agcggcggag gcagcggagg aggaagctgg agccacccgc agttcgagaa atgactcgag
     2701 catgcatcta gaggctggag ccacccgcag ttcgagaaat gactcgagca tgcatctaga
     2761 gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct cgactgtgcc
     2821 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     2881 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     2941 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3001 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     3061 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3121 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3181 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3241 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3301 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3361 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3421 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3481 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     3541 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3601 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3661 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3721 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3781 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3841 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     3901 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3961 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4021 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4081 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     4141 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4201 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4261 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4321 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4381 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4441 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4501 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4561 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4621 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4681 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4741 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4801 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4861 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     4921 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4981 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     5041 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     5101 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5161 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5221 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     5281 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5341 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5401 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5461 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5521 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5581 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5641 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5701 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5761 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5821 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5881 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5941 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6001 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6061 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6121 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6181 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6241 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6301 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6361 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6421 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6481 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6541 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6601 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6661 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6721 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6781 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6841 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6901 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6961 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7021 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7081 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7141 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7201 aagtgccacc tgacgtcgac ggatcggga
//
