LOCUS       sequence-321645-        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        67..370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        371..574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     525..545
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        620..638
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             2887..2913
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2938..3009
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     primer_bind     complement(3062..3081)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3063..3081)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3110..3127)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3116..3323
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3330..3785)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3467..3488)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3679..3698
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3812..3916
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3917..3985,3986..4774)
                     /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(4135..4154)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        4868..5013
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             5056..6093
                     /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(6091..6110)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6135..6183
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6734..7322
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7223..7242
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
       61 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      541 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      601 gcgattacgc caagctcgaa attaaccctc actaaaggga acaaaagctg gctcgaggcc
      661 atgcctggga aactcctctg gggggacatt atggagctgg aagcaccctt ggaggagtcc
      721 gagagccaga agaaggagag gcaaaagagt gacagaagga agtcaaggca ccattatgac
      781 tcggatgaga aatcagaaac aagagaaaat ggtgttacag atgacctgga tgctcccaag
      841 gccaaaaaat ctaaaatgaa agagaagcta aatggagaca ctgaagaagg atttaataga
      901 ctttcagatg aattctccaa atctcataag tcaagaagaa aagatctacc aaatggagat
      961 atagatgaat atgaaaaaaa atcaaagcga gtatcatctt tagatacttc tactcataaa
     1021 tcaagtgata ataaactaga ggagacctta acacgtgaac agaaagaagg agccttctcc
     1081 aattttccta tttctgaaga gactataaag cttctgaaag gtcgaggggt aacatatctc
     1141 tttcctattc aagttaagac ctttggtcct gtatatgaag gaaaagattt aatagctcaa
     1201 gcacggacag gaacaggaaa gacattctct tttgccatcc ccttaattga aagactccaa
     1261 agaaatcaag aaacaattaa aaaaagccgc tcaccaaagg tacttgtttt ggctccaaca
     1321 agggaactgg caaaccaagt agccaaagac ttcaaagata taactaggaa actcagcgtg
     1381 gcgtgttttt atggtggaac atcatatcaa agccaaatta atcatattcg aaatggtatt
     1441 gacatcttgg ttggaacacc tggtcgtatc aaagaccatc tgcagagtgg ccgattggat
     1501 ctttctaaac tgcgacatgt tgtgcttgat gaagtggatc agatgttaga tttaggtttc
     1561 gctgaacaag ttgaagatat tattcatgaa tcctacaaaa ctgattctga agacaatcct
     1621 cagactttac ttttttctgc aacttgccca cagtgggtat acaaagttgc aaaaaaatac
     1681 atgaaatcca gatatgaaca ggttgacctt gttggaaaaa tgactcaaaa ggctgcaact
     1741 actgtggaac atttggccat ccagtgtcat tggtctcaga ggccagcagt tattggagat
     1801 gtccttcaag tctacagtgg gtctgaaggg agggctatta ttttctgtga gaccaagaag
     1861 aatgtaactg aaatggccat gaatccacac ataaaacaga atgcccagtg tttacatggg
     1921 gacattgcac agtcacaaag agaaattaca ctaaaaggct tcagagaagg tagttttaaa
     1981 gttttggtgg caaccaatgt ggctgcccgt ggtttggaca ttcctgaagt tgacctggtg
     2041 attcaaagtt ctcctcctca ggatgttgag tcctatatcc atcgctctgg acgcacaggt
     2101 agagctggac ggacagggat ttgtatatgt ttttatcaac caagagaaag aggtcaacta
     2161 agatatgtgg aacaaaaagc aggaattact tttaaacgtg taggtgttcc ttctacaatg
     2221 gatttagtta aatctaaaag catggatgcc atcaggtctc tggcttccgt ttcttatgct
     2281 gctgttgatt ttttccgacc atcagctcag agactgatag aagagaaagg tgcagtggat
     2341 gcattggctg cagctttagc ccacatttct ggtgcatcaa gctttgaacc acgatctttg
     2401 atcacctctg ataaggggtt tgtgaccatg actctggaaa gcctagagga aatacaggat
     2461 gtcagctgtg cttggaaaga acttaacaga aagctgagta gtaatgcagt gtctcagatt
     2521 accagaatgt gcctcctgaa aggaaatatg ggtgtttgct ttgatgttcc tacaactgag
     2581 tcagaaaggt tacaggcaga gtggcatgat tccgactgga tactctcagt gccagccaaa
     2641 ttacctgaaa ttgaagaata ttatgatgga aacacatctt ctaattccag acagaggagt
     2701 ggctggtcaa gtggtcgatc aggccggtca ggccggtcag gtggtcgatc tggcggccgg
     2761 tcaggtagac agagtcgaca aggaagtcgc tcaggaagtc gacaagatgg tagaagacga
     2821 agtgggaata gaaatcgatc aagaagtggg ggccacaaac ggagttttga cgcggccgca
     2881 atcgattacc catacgatgt tccagattac gctctttagg aattcttcga tgtcgaggat
     2941 tacaaggatg acgacgataa ggactataag gacgatgatg acaaggacta caaagatgat
     3001 gacgataaat agggcccggt accttaatta attaaggtac caggtaagtg tacccaattc
     3061 gccctatagt gagtcgtatt acaattcact cgatcggctc gctgatcagc ctcgactgtg
     3121 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     3181 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     3241 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     3301 gacaatagca ggcatgctgg ggaacgcgta aattgtaagc gttaatattt tgttaaaatt
     3361 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     3421 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     3481 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     3541 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     3601 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     3661 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     3721 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     3781 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3841 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3901 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3961 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4021 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4081 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4141 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4201 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     4261 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4321 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4381 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4441 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     4501 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4561 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4621 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4681 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4741 cgctgagata ggtgcctcac tgattaagca ttggcctcag gattagtgct tcatccccgt
     4801 ggcccgttgc tcgcgtttgc tggcggtgtc cccggaagaa atatatttgc atgtctttag
     4861 ttctatgatg acacaaaccc cgcccagcgt cttgtcattg gcgaattcga acacgcagat
     4921 gcagtcgggg cggcgcggtc cgaggtccac ttcgcatatt aaggtgacgc gtgtggcctc
     4981 gaacaccgag cgaccctgca gcgacccgct taacagcgtc aacagcgtgc cgcagatccc
     5041 ggggggcaat gagatatgaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg
     5101 atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt
     5161 gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat
     5221 ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg
     5281 gaagtgcttg acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca
     5341 cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc
     5401 gcggaggcca tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca
     5461 ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct
     5521 gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg
     5581 caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg
     5641 cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt
     5701 gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg
     5761 aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag
     5821 cttgcaggat cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat
     5881 cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca
     5941 atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc
     6001 gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact
     6061 cgtccggatc gggagatggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     6121 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg
     6181 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     6241 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     6301 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc actggccccg
     6361 tgggttaggg acggggtccc ccatggggaa tggtttatgg ttcgtggggg ttattatttt
     6421 gggcgttgcg tggggtcagg tccacgactg gactgagcag acagacccat ggtttttgga
     6481 tggcctgggc atggaccgca tgtactggcg cgacacgaac accgggcgtc tgtggctgcc
     6541 aaacaccccc gacccccaaa aaccaccgcg cggatttctg gcgccgccgg acgaactaaa
     6601 cctgactacg acatctctgc cccttcttcg ctggtacgag gagcgctttg cctcaggtca
     6661 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6721 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6781 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6841 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6901 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6961 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     7021 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     7081 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     7141 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     7201 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     7261 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     7321 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     7381 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
//
