LOCUS       sequence-84087-e        8249 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for human POP1 with c-terminal 3xFLAG 
            epitopes..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8249)
  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 8249)
  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: pCMVhPOP1_3Flag
FEATURES             Location/Qualifiers
     source          1..8249
                     /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             3757..3828
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     primer_bind     complement(3881..3900)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3882..3900)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3929..3946)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3935..4142
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(4149..4604)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4286..4307)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4498..4517
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4631..4735
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4736..4804,4805..5593)
                     /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(4954..4973)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        5687..5832
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             5875..6912
                     /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(6910..6929)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6954..7002
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7553..8141
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8042..8061
                     /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 gctcgacgcc
      661 accatgtcaa atgcaaaaga aagaaaacac gccaagaaaa tgagaaacca gcctaccaat
      721 gtgactctgt cctctggctt tgtggctgac agaggtgtaa agcaccacag tggaggtgaa
      781 aaacctttcc aagctcaaaa acaagagcct catcctggaa cttcacgaca gcggcaaacc
      841 agagtcaacc cccattctct gcctgaccct gaagtgaatg agcagtcttc ctccaaaggg
      901 atgtttagaa aaaagggagg atggaaagca ggtcccgagg gcacgtctca ggagatcccc
      961 aagtatataa ctgcttctac ttttgctcaa gcacgagctg ctgaaatcag tgctatgtta
     1021 aaagctgtga cccagaagtc ttcgaattca ctggtttttc agactctgcc acggcacatg
     1081 cgacgaagag ccatgagcca caacgtcaaa cgccttccca gacggttaca ggagattgcc
     1141 cagaaagagg cggagaaagc cgtacatcag aaaaaagaac attcaaaaaa taaatgccat
     1201 aaagctcgaa gatgtcacat gaaccggacg ctagaattta accgtagaca aaagaagaac
     1261 atttggttag aaactcacat ctggcacgcc aagcggtttc atatggtcaa gaagtggggc
     1321 tactgccttg gggagaggcc aacagtcaag agccacagag cctgctatcg agccatgacg
     1381 aaccggtgcc tcctgcagga tttatcctat tactgttgtt tggagttgaa aggcaaagag
     1441 gaagaaatac taaaggcgct ttctggaatg tgtaacatag acacagggct gacgtttgca
     1501 gcagttcact gcttgtctgg aaagcgccaa gggagccttg tgctttatcg ggtgaataaa
     1561 tatcccagag aaatgcttgg gcctgttacg tttatctgga agtcccagag gaccccgggt
     1621 gacccttctg agagcaggca gctgtggatc tggctgcatc caacccttaa acaggatatc
     1681 ttagaggaaa taaaagcagc gtgccagtgt gtggaaccca tcaaatcagc tgtctgcatc
     1741 gctgacccac ttccaacacc atcccaagaa aaaagccaaa ctgaattgcc tgacgagaaa
     1801 attggcaaga aaagaaaaag gaaagatgat ggagaaaatg ctaaaccaat taaaaaaatt
     1861 atcggtgatg gaactagaga tccatgtcta ccatactctt ggatctctcc aaccacaggc
     1921 attataatca gcgatttgac gatggagatg aacagattcc ggctgattgg gccactttcc
     1981 cactccatcc taactgaagc aataaaagct gcttctgtcc acactgtggg agaggacaca
     2041 gaggagacac ctcaccgctg gtggatagaa acctgtaaga aacctgacag cgtttccctt
     2101 cattgcagac aagaagccat tttcgagttg ttgggaggaa taacatcacc agcagaaatt
     2161 ccggcaggta ctattctggg actgacagtt ggggatcctc gaataaattt gccccaaaag
     2221 aagtccaacg ctttgcccaa tccagaaaaa tgccaagata atgagaaagt tagacagctg
     2281 cttctggagg gtgtgcctgt ggaatgtacg catagcttta tctggaacca agatatctgt
     2341 aagagtgtca cagagaataa aatctcggat caggatttaa accggatgag gagtgaattg
     2401 ctggtgcctg ggtcacagct tattttaggt ccccatgaat ccaagatacc tatacttttg
     2461 attcagcagc caggaaaagt gactggtgaa gatcgactag gctggggaag tggctgggat
     2521 gtcctactcc caaagggctg gggcatggct ttctggattc catttattta tcgaggtgtg
     2581 agagtcggag ggttgaaaga gtctgcagtg cattctcagt ataagaggtc gcctaatgtc
     2641 ccaggcgatt ttccagactg ccctgccggg atgctgtttg cggaagagca agctaagaat
     2701 cttcttgaaa agtacaaaag acgccctcct gcaaaacggc ccaactacgt taagcttggc
     2761 actctggcac ctttctgctg tccctgggag cagttaactc aagactggga gtcaagagtc
     2821 caggcttacg aagaaccttc tgtagcttca tctccaaatg gtaaggagag tgacctaaga
     2881 agatctgagg tgccttgtgc tcccatgcct aaaaaaactc atcagccatc tgatgaagtg
     2941 ggcacatcca tagagcaccc cagggaggca gaggaggtaa tggatgcagg gtgtcaagaa
     3001 tcggcagggc ctgagaggat cacagaccag gaggccagtg aaaaccatgt tgctgccaca
     3061 gggagtcacc tctgcgttct caggagtaga aaattactga agcaactgtc agcctggtgt
     3121 gggcccagtt ctgaggatag tcggggaggc cggcgagctc ccggcagagg ccagcaagga
     3181 ttgaccagag aggcttgcct gtccatcttg ggccacttcc ccagggccct ggtttgggtc
     3241 agcctgtccc tgctcagcaa gggcagcccc gagcctcaca ccatgatctg tgtcccagcc
     3301 aaggaggact tcctccagct ccatgaggac tggcattact gtgggcccca ggaatccaaa
     3361 cacagtgacc cattcaggag caagatcctg aaacagaaag agaagaagaa aagggagaag
     3421 aggcagaagc caggacgtgc ctcttctgat ggcccggcgg gggaagagcc cgtggctggg
     3481 caggaagctc tgactctagg gctgtggtca ggccctctgc cgcgtgtgac gttgcactgc
     3541 tccagaactc tcctaggctt tgtgactcag ggagattttt ccatggctgt tggctgtgga
     3601 gaagccctgg ggtttgttag cttgacaggc ttgctggata tggtgtccag ccagcctgca
     3661 gcgcagaggg gcttagtgct actgaggcct cccgcctctc tgcagtatcg atttgcgagg
     3721 attgctattg aggtggcggc cgcaatcgat gtcgaggatt acaaggatga cgacgataag
     3781 gactataagg acgatgatga caaggactac aaagatgatg acgataaata gggcccggta
     3841 ccttaattaa ttaaggtacc aggtaagtgt acccaattcg ccctatagtg agtcgtatta
     3901 caattcactc gatcggctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     3961 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     4021 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     4081 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     4141 gaacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     4201 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     4261 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     4321 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     4381 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     4441 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     4501 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     4561 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     4621 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4681 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4741 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4801 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4861 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4921 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4981 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5041 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5101 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5161 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5221 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5281 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5341 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5401 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5461 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5521 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5581 gattaagcat tggcctcagg attagtgctt catccccgtg gcccgttgct cgcgtttgct
     5641 ggcggtgtcc ccggaagaaa tatatttgca tgtctttagt tctatgatga cacaaacccc
     5701 gcccagcgtc ttgtcattgg cgaattcgaa cacgcagatg cagtcggggc ggcgcggtcc
     5761 gaggtccact tcgcatatta aggtgacgcg tgtggcctcg aacaccgagc gaccctgcag
     5821 cgacccgctt aacagcgtca acagcgtgcc gcagatcccg gggggcaatg agatatgaaa
     5881 aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc
     5941 tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga
     6001 gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat
     6061 gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggaa
     6121 ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac
     6181 ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggccat ggatgcgatc
     6241 gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt
     6301 caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg
     6361 caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg
     6421 ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac
     6481 aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc
     6541 ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg
     6601 gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc gccgcggctc
     6661 cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat
     6721 ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg
     6781 actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta
     6841 gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccggatcg ggagatgggg
     6901 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     6961 aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     7021 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     7081 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     7141 acgtcggggc ggcaggccct gccatagcca ctggccccgt gggttaggga cggggtcccc
     7201 catggggaat ggtttatggt tcgtgggggt tattattttg ggcgttgcgt ggggtcaggt
     7261 ccacgactgg actgagcaga cagacccatg gtttttggat ggcctgggca tggaccgcat
     7321 gtactggcgc gacacgaaca ccgggcgtct gtggctgcca aacacccccg acccccaaaa
     7381 accaccgcgc ggatttctgg cgccgccgga cgaactaaac ctgactacga catctctgcc
     7441 ccttcttcgc tggtacgagg agcgctttgc ctcaggtcat gaccaaaatc ccttaacgtg
     7501 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     7561 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     7621 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     7681 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     7741 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     7801 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     7861 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     7921 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     7981 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     8041 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     8101 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     8161 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     8221 ctgattctgt ggataaccgt attaccgcc
//
