LOCUS       sequence-327029-        6869 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human HP1BP3 mammalian expression vector with C-terminal HA tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6869)
  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 6869)
  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-hHP1BP3-HA
FEATURES             Location/Qualifiers
     source          1..6869
                     /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             2260..2286
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2311..2382
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     primer_bind     complement(2435..2454)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2436..2454)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2483..2500)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2489..2696
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2703..3158)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2840..2861)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3052..3071
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3185..3289
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3290..3358,3359..4147)
                     /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(3508..3527)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        4241..4386
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             4429..5466
                     /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(5464..5483)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5508..5556
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6107..6695
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6596..6615
                     /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 gaccatggcg
      601 actgatacgt ctcaaggtga actcgtccat cctaaggcac tcccacttat agtaggagct
      661 cagctgatcc acgcggacaa gaaggtagaa gatagcacca tgccgattcg tcgaactgtg
      721 aattctaccc gggaaactcc tcccaaaagc aagcttgctg aaggggagga agaaaagcca
      781 gaaccagaca taagttcaga ggaatctgtc tccactgtag aagaacaaga gaatgaaact
      841 ccacctgcta cttcgagtga ggcagagcag ccaaaggggg aacctgagaa tgaagagaag
      901 gaagaaaata agtcttctga ggaaaccaaa aaggatgaga aagatcagtc taaagaaaag
      961 gagaagaaag tgaaaaaaac aattccttcc tgggctaccc tttctgccag ccagctagcc
     1021 agggcccaga aacaaacacc gatggcttct tccccacgtc ccaagatgga tgcaatctta
     1081 actgaggcca ttaaggcatg cttccagaag agtggtgcat cagtggttgc tattcgaaaa
     1141 tacatcatcc ataagtatcc ttctctggag ctggagagaa ggggttatct ccttaaacaa
     1201 gcactgaaaa gagaattaaa tagaggagtc atcaaacagg ttaaaggaaa aggtgcttct
     1261 ggaagttttg ttgtggttca gaaatcaaga aaaacacctc agaaatccag aaacagaaag
     1321 aataggagct ctgcagtgga tccagaacca caagtaaaat tggaggatgt cctcccactg
     1381 gcctttactc gcctttgtga acctaaagaa gcttcctaca gtctcatcag gaaatatgtg
     1441 tctcagtatt atcctaagct tagagtggac atcaggcctc agctgttgaa gaacgctctg
     1501 cagagagcag tagagagggg ccagttagaa cagataactg gcaaaggtgc ttcggggaca
     1561 ttccagctga agaaatcagg ggagaaaccc ctgcttggtg gaagcctgat ggaatatgca
     1621 atcttgtctg ccattgctgc catgaatgag ccgaagacct gctctactac tgctctgaag
     1681 aagtatgtcc tagagaatca cccaggaacc aattctaact atcaaatgca tttgctgaaa
     1741 aaaaccctgc agaaatgcga aaagaatggg tggatggaac agatctctgg gaaagggttc
     1801 agtggcacct tccagctctg ttttccctat tatcccagcc caggagttct gtttccgaag
     1861 aaagagccag atgattctag agatgaggat gaagatgaag atgagtcatc agaagaagac
     1921 tctgaggatg aagagccgcc acctaagaga aggttgcaga agaaaacccc agccaagtcc
     1981 ccagggaagg ccgcatctgt gaagcagaga gggtccaaac ctgcacctaa agtctcagct
     2041 gcccagcggg ggaaagctag gcccttgcct aagaaagcac ctcctaaggc caaaacgcct
     2101 gccaagaaga ccagaccttc atccacagtc atcaagaaac ctagtggtgg ctcctcaaag
     2161 aagcctgcaa ccagtgcaag aaaggaagta aaattgccgg gcaagggcaa atccaccatg
     2221 aagaagtctt tcagagtgaa aaaggcggcc gcaatcgatt acccatacga tgttccagat
     2281 tacgctcttt aggaattctt cgatgtcgag gattacaagg atgacgacga taaggactat
     2341 aaggacgatg atgacaagga ctacaaagat gatgacgata aatagggccc ggtaccttaa
     2401 ttaattaagg taccaggtaa gtgtacccaa ttcgccctat agtgagtcgt attacaattc
     2461 actcgatcgg ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     2521 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2581 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2641 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggaacgc
     2701 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     2761 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     2821 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     2881 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     2941 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     3001 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     3061 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     3121 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     3181 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3241 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     3301 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3361 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     3421 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     3481 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     3541 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     3601 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     3661 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     3721 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     3781 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     3841 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     3901 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     3961 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4021 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4081 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4141 gcattggcct caggattagt gcttcatccc cgtggcccgt tgctcgcgtt tgctggcggt
     4201 gtccccggaa gaaatatatt tgcatgtctt tagttctatg atgacacaaa ccccgcccag
     4261 cgtcttgtca ttggcgaatt cgaacacgca gatgcagtcg gggcggcgcg gtccgaggtc
     4321 cacttcgcat attaaggtga cgcgtgtggc ctcgaacacc gagcgaccct gcagcgaccc
     4381 gcttaacagc gtcaacagcg tgccgcagat cccggggggc aatgagatat gaaaaagcct
     4441 gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac
     4501 ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt
     4561 ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat
     4621 cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg ggaattcagc
     4681 gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca agacctgcct
     4741 gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg
     4801 gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat cggtcaatac
     4861 actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact
     4921 gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg
     4981 gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc
     5041 ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat
     5101 tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag
     5161 cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg
     5221 tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat
     5281 gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc
     5341 gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta
     5401 ctcgccgata gtggaaaccg acgccccagc actcgtccgg atcgggagat gggggaggct
     5461 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     5521 acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     5581 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     5641 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     5701 gggcggcagg ccctgccata gccactggcc ccgtgggtta gggacggggt cccccatggg
     5761 gaatggttta tggttcgtgg gggttattat tttgggcgtt gcgtggggtc aggtccacga
     5821 ctggactgag cagacagacc catggttttt ggatggcctg ggcatggacc gcatgtactg
     5881 gcgcgacacg aacaccgggc gtctgtggct gccaaacacc cccgaccccc aaaaaccacc
     5941 gcgcggattt ctggcgccgc cggacgaact aaacctgact acgacatctc tgccccttct
     6001 tcgctggtac gaggagcgct ttgcctcagg tcatgaccaa aatcccttaa cgtgagtttt
     6061 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     6121 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     6181 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     6241 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     6301 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     6361 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6421 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6481 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     6541 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6601 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6661 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6721 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6781 ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca
     6841 ttagttcata gcccatatat ggagttccg
//
