LOCUS       sequence-327814-        6509 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human RBM34 mammalian expression vector with C-terminal HA tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6509)
  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 6509)
  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-hRBM34-HA
FEATURES             Location/Qualifiers
     source          1..6509
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        554..572
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             1900..1926
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1951..2022
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     primer_bind     complement(2075..2094)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2076..2094)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2123..2140)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2129..2336
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2343..2798)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2480..2501)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2692..2711
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2825..2929
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2930..2998,2999..3787)
                     /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(3148..3167)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        3881..4026
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             4069..5106
                     /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(5104..5123)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5148..5196
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5747..6335
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6236..6255
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgatt
      541 acgccaagct cgaaattaac cctcactaaa gggaacaaaa gctggctcga gaccatggcc
      601 ttggaaggga tgagcaaacg gaagagaaag agaagtgtcc aggagggaga gaatcctgac
      661 gacggcgttc gcgggagtcc gccggaagac tacaggcttg gacaggtcgc cagtagctta
      721 tttcgcggcg aacaccattc cagaggtggc accggtcggc tggcgtccct cttcagttct
      781 ctggagcccc agattcaacc cgtgtacgtg cctgtgccta aacaaaccat caaaaaaacg
      841 aaacggaatg aggaggaaga aagtacatcc cagattgaaa gaccactttc gcaagaacct
      901 gccaaaaaag tgaaagcgaa gaagaaacac actaacgcag aaaaaaagtt ggcagacagg
      961 gaaagcgctc tagcgagtgc tgatttagaa gaagaaattc accagaaaca agggcagaaa
     1021 aggaaaaatt ctcaacctgg tgttaaagta gcagatagaa aaatacttga tgacacagaa
     1081 gacacagttg tcagtcaaag aaagaaaatt caaatcaacc aagaagaaga gagattaaag
     1141 aatgagagaa ctgtgtttgt tgggaatttg cctgttacat gtaataagaa gaagctgaag
     1201 tcgtttttta aagagtatgg acaaatagaa tctgtacgat ttcgttctct gattccagca
     1261 gagggaacgc tatccaaaaa gttggcagca ataaaacgta aaattcatcc tgatcagaaa
     1321 aatattaatg cctatgttgt gtttaaggag gagagtgctg ccacgcaagc attgaaaaga
     1381 aatggggccc agattgcaga tggatttcgt attagagttg atctcgcatc tgagacctca
     1441 tctagagaca agagatcggt ttttgtgggg aatctccctt ataaagttga agaatctgcc
     1501 attgagaagc actttctgga ctgtggaagt atcatggccg tgaggattgt gagagacaaa
     1561 atgacaggca tcggcaaagg gtttggctat gtgctctttg agaatacaga ttctgttcat
     1621 cttgctctga aattaaataa ttctgaactc atggggagaa aactcagagt catgcgttct
     1681 gttaataaag aaaaatttaa acaacaaaat tcaaatccac gattgaagaa tgtcagtaaa
     1741 cctaagcagg gacttaattt tacttccaaa actgcagaag gacatcctaa aagcttattt
     1801 attggagaaa aagctgttct ccttaaaacg aagaagaaag gacagaagaa aagtggacgc
     1861 cctaagaaac agagaaaaca gaaagcggcc gcaatcgatt acccatacga tgttccagat
     1921 tacgctcttt aggaattctt cgatgtcgag gattacaagg atgacgacga taaggactat
     1981 aaggacgatg atgacaagga ctacaaagat gatgacgata aatagggccc ggtaccttaa
     2041 ttaattaagg taccaggtaa gtgtacccaa ttcgccctat agtgagtcgt attacaattc
     2101 actcgatcgg ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     2161 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2221 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2281 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggaacgc
     2341 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     2401 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     2461 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     2521 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     2581 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     2641 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     2701 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     2761 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     2821 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2881 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     2941 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3001 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     3061 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     3121 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     3181 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     3241 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     3301 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     3361 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     3421 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     3481 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     3541 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     3601 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     3661 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     3721 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     3781 gcattggcct caggattagt gcttcatccc cgtggcccgt tgctcgcgtt tgctggcggt
     3841 gtccccggaa gaaatatatt tgcatgtctt tagttctatg atgacacaaa ccccgcccag
     3901 cgtcttgtca ttggcgaatt cgaacacgca gatgcagtcg gggcggcgcg gtccgaggtc
     3961 cacttcgcat attaaggtga cgcgtgtggc ctcgaacacc gagcgaccct gcagcgaccc
     4021 gcttaacagc gtcaacagcg tgccgcagat cccggggggc aatgagatat gaaaaagcct
     4081 gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac
     4141 ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt
     4201 ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat
     4261 cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg ggaattcagc
     4321 gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca agacctgcct
     4381 gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg
     4441 gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat cggtcaatac
     4501 actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact
     4561 gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg
     4621 gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc
     4681 ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat
     4741 tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag
     4801 cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg
     4861 tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat
     4921 gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc
     4981 gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta
     5041 ctcgccgata gtggaaaccg acgccccagc actcgtccgg atcgggagat gggggaggct
     5101 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     5161 acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     5221 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     5281 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     5341 gggcggcagg ccctgccata gccactggcc ccgtgggtta gggacggggt cccccatggg
     5401 gaatggttta tggttcgtgg gggttattat tttgggcgtt gcgtggggtc aggtccacga
     5461 ctggactgag cagacagacc catggttttt ggatggcctg ggcatggacc gcatgtactg
     5521 gcgcgacacg aacaccgggc gtctgtggct gccaaacacc cccgaccccc aaaaaccacc
     5581 gcgcggattt ctggcgccgc cggacgaact aaacctgact acgacatctc tgccccttct
     5641 tcgctggtac gaggagcgct ttgcctcagg tcatgaccaa aatcccttaa cgtgagtttt
     5701 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5761 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5821 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5881 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5941 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     6001 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6061 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6121 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     6181 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6241 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6301 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6361 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6421 ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca
     6481 ttagttcata gcccatatat ggagttccg
//
