LOCUS       sequence-327030-        7430 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human DDX50 mammalian expression vector with C-terminal HA tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7430)
  AUTHORS   Li S, Wang L, Fu B, Berman MA, Diallo A, Dorf ME
  TITLE     TRIM65 regulates microRNA activity by ubiquitination of TNRC6.
  JOURNAL   Proc Natl Acad Sci U S A. 2014 May 13;111(19):6970-5. doi: 
            10.1073/pnas.1322545111. Epub 2014 Apr 28.
  PUBMED    24778252
REFERENCE   2  (bases 1 to 7430)
  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"; doi: 
            "10.1073/pnas.1322545111"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2014-05-13- 13"; volume: "111"; issue: "19"; pages: "6970-5"
COMMENT     Sequence Label: pCMV3Tag8li-hDDX50-HA
FEATURES             Location/Qualifiers
     source          1..7430
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        121..424
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        425..628
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     579..599
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        674..692
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             2941..2967
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2992..3063
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     primer_bind     complement(3116..3135)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3117..3135)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3164..3181)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3170..3377
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3384..3839)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3521..3542)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3733..3752
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3866..3970
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3971..4039,4040..4828)
                     /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(4189..4208)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        4922..5067
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             5110..6147
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDRE
                     MGEAN"
     primer_bind     complement(6145..6164)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6189..6237
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6788..7376
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7277..7296
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
       61 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      121 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      181 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      241 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      301 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      361 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      421 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      481 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      541 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      601 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgatt
      661 acgccaagct cgaaattaac cctcactaaa gggaacaaaa gctggctcga ggccatgcct
      721 gggaaactcc tctgggggga cattatggag ctggaagcac ccttggagga gtccgagagc
      781 cagaagaagg agaggcaaaa gagtgacaga aggaagtcaa ggcaccatta tgactcggat
      841 gagaaatcag aaacaagaga aaatggtgtt acagatgacc tggatgctcc caaggccaaa
      901 aaatctaaaa tgaaagagaa gctaaatgga gacactgaag aaggatttaa tagactttca
      961 gatgaattct ccaaatctca taagtcaaga agaaaagatc taccaaatgg agatatagat
     1021 gaatatgaaa aaaaatcaaa gcgagtatca tctttagata cttctactca taaatcaagt
     1081 gataataaac tagaggagac cttaacacgt gaacagaaag aaggagcctt ctccaatttt
     1141 cctatttctg aagagactat aaagcttctg aaaggtcgag gggtaacata tctctttcct
     1201 attcaagtta agacctttgg tcctgtatat gaaggaaaag atttaatagc tcaagcacgg
     1261 acaggaacag gaaagacatt ctcttttgcc atccccttaa ttgaaagact ccaaagaaat
     1321 caagaaacaa ttaaaaaaag ccgctcacca aaggtacttg ttttggctcc aacaagggaa
     1381 ctggcaaacc aagtagccaa agacttcaaa gatataacta ggaaactcag cgtggcgtgt
     1441 ttttatggtg gaacatcata tcaaagccaa attaatcata ttcgaaatgg tattgacatc
     1501 ttggttggaa cacctggtcg tatcaaagac catctgcaga gtggccgatt ggatctttct
     1561 aaactgcgac atgttgtgct tgatgaagtg gatcagatgt tagatttagg tttcgctgaa
     1621 caagttgaag atattattca tgaatcctac aaaactgatt ctgaagacaa tcctcagact
     1681 ttactttttt ctgcaacttg cccacagtgg gtatacaaag ttgcaaaaaa atacatgaaa
     1741 tccagatatg aacaggttga ccttgttgga aaaatgactc aaaaggctgc aactactgtg
     1801 gaacatttgg ccatccagtg tcattggtct cagaggccag cagttattgg agatgtcctt
     1861 caagtctaca gtgggtctga agggagggct attattttct gtgagaccaa gaagaatgta
     1921 actgaaatgg ccatgaatcc acacataaaa cagaatgccc agtgtttaca tggggacatt
     1981 gcacagtcac aaagagaaat tacactaaaa ggcttcagag aaggtagttt taaagttttg
     2041 gtggcaacca atgtggctgc ccgtggtttg gacattcctg aagttgacct ggtgattcaa
     2101 agttctcctc ctcaggatgt tgagtcctat atccatcgct ctggacgcac aggtagagct
     2161 ggacggacag ggatttgtat atgtttttat caaccaagag aaagaggtca actaagatat
     2221 gtggaacaaa aagcaggaat tacttttaaa cgtgtaggtg ttccttctac aatggattta
     2281 gttaaatcta aaagcatgga tgccatcagg tctctggctt ccgtttctta tgctgctgtt
     2341 gattttttcc gaccatcagc tcagagactg atagaagaga aaggtgcagt ggatgcattg
     2401 gctgcagctt tagcccacat ttctggtgca tcaagctttg aaccacgatc tttgatcacc
     2461 tctgataagg ggtttgtgac catgactctg gaaagcctag aggaaataca ggatgtcagc
     2521 tgtgcttgga aagaacttaa cagaaagctg agtagtaatg cagtgtctca gattaccaga
     2581 atgtgcctcc tgaaaggaaa tatgggtgtt tgctttgatg ttcctacaac tgagtcagaa
     2641 aggttacagg cagagtggca tgattccgac tggatactct cagtgccagc caaattacct
     2701 gaaattgaag aatattatga tggaaacaca tcttctaatt ccagacagag gagtggctgg
     2761 tcaagtggtc gatcaggccg gtcaggccgg tcaggtggtc gatctggcgg ccggtcaggt
     2821 agacagagtc gacaaggaag tcgctcagga agtcgacaag atggtagaag acgaagtggg
     2881 aatagaaatc gatcaagaag tgggggccac aaacggagtt ttgacgcggc cgcaatcgat
     2941 tacccatacg atgttccaga ttacgctctt taggaattct tcgatgtcga ggattacaag
     3001 gatgacgacg ataaggacta taaggacgat gatgacaagg actacaaaga tgatgacgat
     3061 aaatagggcc cggtacctta attaattaag gtaccaggta agtgtaccca attcgcccta
     3121 tagtgagtcg tattacaatt cactcgatcg gctcgctgat cagcctcgac tgtgccttct
     3181 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     3241 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3301 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3361 agcaggcatg ctggggaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     3421 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     3481 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     3541 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     3601 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     3661 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     3721 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     3781 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     3841 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3901 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3961 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4021 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4081 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4141 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4201 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4261 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     4321 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4381 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     4441 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     4501 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     4561 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     4621 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     4681 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     4741 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     4801 gataggtgcc tcactgatta agcattggcc tcaggattag tgcttcatcc ccgtggcccg
     4861 ttgctcgcgt ttgctggcgg tgtccccgga agaaatatat ttgcatgtct ttagttctat
     4921 gatgacacaa accccgccca gcgtcttgtc attggcgaat tcgaacacgc agatgcagtc
     4981 ggggcggcgc ggtccgaggt ccacttcgca tattaaggtg acgcgtgtgg cctcgaacac
     5041 cgagcgaccc tgcagcgacc cgcttaacag cgtcaacagc gtgccgcaga tcccgggggg
     5101 caatgagata tgaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa
     5161 aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc
     5221 agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc
     5281 tacaaagatc gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg
     5341 cttgacattg gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt
     5401 gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag
     5461 gccatggatg cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga
     5521 ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc
     5581 catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct
     5641 ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg
     5701 gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg
     5761 agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg
     5821 tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca
     5881 ggatcgccgc ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc
     5941 ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc
     6001 cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg
     6061 accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg
     6121 gatcgggaga tgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     6181 cgctatgacg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata
     6241 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     6301 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     6361 cagggctcgc agccaacgtc ggggcggcag gccctgccat agccactggc cccgtgggtt
     6421 agggacgggg tcccccatgg ggaatggttt atggttcgtg ggggttatta ttttgggcgt
     6481 tgcgtggggt caggtccacg actggactga gcagacagac ccatggtttt tggatggcct
     6541 gggcatggac cgcatgtact ggcgcgacac gaacaccggg cgtctgtggc tgccaaacac
     6601 ccccgacccc caaaaaccac cgcgcggatt tctggcgccg ccggacgaac taaacctgac
     6661 tacgacatct ctgccccttc ttcgctggta cgaggagcgc tttgcctcag gtcatgacca
     6721 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6781 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6841 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6901 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     6961 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7021 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7081 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7141 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7201 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7261 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7321 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7381 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
//
