LOCUS       sequence-318524-        5598 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses murine Ddx58 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5598)
  AUTHORS   Gstalder C, Liu D, Miao D, Lutterbach B, DeVine AL, Lin C, Shettigar
            M, Pancholi P, Buchbinder EI, Carter SL, Manos MP, Rojas-Rudilla V, 
            Brennick R, Gjini E, Chen PH, Lako A, Rodig S, Yoon CH, Freeman GJ, 
            Barbie DA, Hodi FS, Miles W, Van Allen EM, Haq R
  TITLE     Inactivation of Fbxw7 impairs dsRNA sensing and confers resistance 
            to PD-1 blockade.
  JOURNAL   Cancer Discov. 2020 May 5. pii: 2159-8290.CD-19-1416. doi: 
            10.1158/2159-8290.CD-19-1416.
  PUBMED    32371478
REFERENCE   2  (bases 1 to 5598)
  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: "Cancer 
            Discov. 2020 May 5. pii: 2159-8290.CD-19-1416. doi: 
            10.1158/2159-8290.CD-19-1416."
COMMENT     Sequence Label: pENTR223.1_mmDdx58 (closed)
FEATURES             Location/Qualifiers
     source          1..5598
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3477..3576)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3593..3612)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3594..3612)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3617..3633)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3617..3633)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4064..4855
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      4948..5536
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5437..5456
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcagaaggg ccgtcaaggc ccaccatgac agcggagcag cggcagaatc tgcaagcatt
      721 cagagactat atcaagaaga ttctggaccc cacctacatc ctcagctaca tgagttcctg
      781 gctcgaggat gaggaggtgc agtacattca ggctgagaag aacaacaagg gcccaatgga
      841 agctgcctca ctcttcctcc agtacctgtt gaagctgcag tcagagggct ggttccaggc
      901 ctttttggat gccctgtacc atgcaggtta ctgtggactt tgtgaagcca tcgaaagttg
      961 ggactttcaa aaaattgaaa agttagagga acacagatta cttttaagac gtttagaacc
     1021 agaatttaag gccacagttg atccaaatga tatcctttct gaactatccg aatgtttgat
     1081 taatcaggaa tgtgaagaaa tcagacagat ccgagacact aaagggagaa tggcaggtgc
     1141 ggagaagatg gccgaatgtc ttatcagatc cgacaaggaa aactggccaa aggtcttgca
     1201 acttgctttg gagaaagaca acagcaagtt tagtgaattg tggattgttg ataaaggttt
     1261 caaaagggct gaaagcaagg ctgatgagga tgatggagcg gaggcgtcca gcatccagat
     1321 tttcattcag gaagagccag agtgtcagaa tctcagtcag aatcccgggc ctccttcaga
     1381 agcgtcttct aataatttac acagcccatt gaaaccaaga aattaccaac tggagcttgc
     1441 cctgcctgcc aagaaaggga aaaatacaat aatatgtgcc cctactggtt gtggaaaaac
     1501 ctttgtgtcg cttcttatat gtgaacacca tcttaaaaaa ttcccatgtg gacaaaaagg
     1561 gaaagtggtc ttcttcgcta accaaattcc tgtctatgag cagcaggcaa ctgtgttctc
     1621 acgatatttt gaaagacttg ggtacaacat tgcgagcatt tctggggcaa catctgatag
     1681 cgtctcagtg cagcacatca ttgaagacaa tgatatcatc atcctgacac cccagattct
     1741 tgtgaacaat ctcaacaacg gagccatccc ctcgttgtct gtcttcactc tgatgatatt
     1801 tgatgagtgt cataacacta gcaaaaacca cccatacaat cagatcatgt tcagatacct
     1861 agaccacaaa cttggagagt cacgggaccc actgcctcag gtcgttgggc tgactgcctc
     1921 cgtcggcgtt ggagatgcta agaccgcgga ggaagccatg caacatatct gtaaactctg
     1981 tgccgccctg gatgcctccg tgattgccac agtcagagac aacgttgcag aactggaaca
     2041 ggtcgtttat aagccccaga aaatttccag gaaagtggca tcccggactt cgaacacgtt
     2101 taaatgcatc atctctcagc tgatgaagga gacagagaag ctagccaagg atgtctccga
     2161 ggaacttgga aagctttttc aaattcaaaa cagagaattc ggcacccaga aatatgaaca
     2221 gtggattgtc ggcgtccaca aagcgtgctc agtgtttcag atggcagaca aagaggagga
     2281 gagccgggtc tgcaaagcgc tcttcctgta cacatcacat ttgcggaaat acaacgatgc
     2341 actcatcatc agtgaggatg cacagatgac agacgctcta aattacctca aagccttctt
     2401 ccacgatgtc cgagaagcag cattcgatga gaccgagcga gagcttactc ggaggtttga
     2461 agaaaaacta gaggaattag aaaaagtttc cagggatccc agcaatgaga atcctaaact
     2521 aagagacctc tacttggtct tacaagaaga gtaccactta aagccagaga ccaagaccat
     2581 tctcttcgtg aagaccagag cactcgtgga tgctctgaag aaatggattg aagaaaatcc
     2641 tgcactaagc tttctaaagc ctggcatact gactgggcgt ggcagaacaa accgggcaac
     2701 aggaatgacg ctcccggcac agaagtgtgt gctggaggca ttcagagcca gcggagataa
     2761 caatattctg attgctacct cggtcgctga tgaaggcatt gacattgctg agtgcaatct
     2821 cgtcattctc tatgagtacg tgggcaacgt catcaagatg atccaaacca gaggccgagg
     2881 aagagcacga gatagcaagt gcttcctcct gaccagcagc gctgacgtga ttgaaaaaga
     2941 aaaggcgaac atgatcaagg aaaaaataat gaatgaatcc atcttaagac tgcagacatg
     3001 ggatgaaatg aaatttggaa agacggttca ccgcatacag gtgaatgaaa aactcctcag
     3061 agacagtcag cacaaaccac aacctgttcc tgacaaagaa aacaagaaac tgctgtgtgg
     3121 aaagtgcaag aattttgcgt gctacacagc tgacattcga gtggttgaga cgtcccacta
     3181 cactgtcctt ggagacgctt ttaaggagcg ctttgtgtgt aagccacacc ctaaaccaaa
     3241 gatctatgac aattttgaga agaaagcaaa gatattctgc gccaaacaga actgtagcca
     3301 cgactgggga atttttgtga gatacaagac gttcgagatt ccagtcataa aaattgaaag
     3361 tttcgtcgtg gaagatattg tgagcggagt tcagaaccgg cactcaaagt ggaaggactt
     3421 tcattttgaa aggatacagt tcgatcctgc agaaatgtcc gtatagggcc tcatgggccc
     3481 agctttcttg tacaaagttg gcattataaa aaataattgc tcatcaattt gttgcaacga
     3541 acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag
     3601 tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct
     3661 ctgatgttac attgcacaag ataaaaatat atcatcatgc ctcctctaga ccagccagga
     3721 cagaaatgcc tcgacttcgc tgctacccaa ggttgccggg tgacgcacac cgtggaaacg
     3781 gatgaaggca cgaacccagt ggacataagc ctgttcggtt cgtaagctgt aatgcaagta
     3841 gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc
     3901 agtggcggtt ttcatggctt gttatgactg tttttttggg gtacagtcta tgcctcgggc
     3961 atccaagcag caagcgcgtt acgccgtggg tcgatgtttg atgttatgga gcagcaacga
     4021 tgttacgcag cagggcagtc gccctaaaac aaagttaaac attatgaggg aagcggtgat
     4081 cgccgaagta tcgactcaac tatcagaggt agttggcgtc atcgagcgcc atctcgaacc
     4141 gacgttgctg gccgtacatt tgtacggctc cgcagtggat ggcggcctga agccacacag
     4201 tgatattgat ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt
     4261 gatcaacgac cttttggaaa cttcggcttc ccctggagag agcgagattc tccgcgctgt
     4321 agaagtcacc attgttgtgc acgacgacat cattccgtgg cgttatccag ctaagcgcga
     4381 actgcaattt ggagaatggc agcgcaatga cattcttgca ggtatcttcg agccagccac
     4441 gatcgacatt gatctggcta tcttgctgac aaaagcaaga gaacatagcg ttgccttggt
     4501 aggtccagcg gcggaggaac tctttgatcc ggttcctgaa caggatctat ttgaggcgct
     4561 aaatgaaacc ttaacgctat ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt
     4621 agtgcttacg ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga
     4681 tgtcgctgcc gactgggcaa tggagcgcct gccggcccag tatcagcccg tcatacttga
     4741 agctagacag gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt
     4801 ggaagaattt gtccactacg tgaaaggcga gatcaccaag gtagtcggca aataaccctc
     4861 gagccaccca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     4921 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4981 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5041 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5101 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5161 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5221 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5281 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5341 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5401 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5461 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5521 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5581 gccttttgct cacatgtt
//
