LOCUS       sequence-180668-        8763 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses V5-ANKRD52.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8763)
  AUTHORS   Golden RJ, Chen B, Li T, Braun J, Manjunath H, Chen X, Wu J, Schmid 
            V, Chang TC, Kopp F, Ramirez-Martinez A, Tagliabracci VS, Chen ZJ, 
            Xie Y, Mendell JT
  TITLE     An Argonaute phosphorylation cycle promotes microRNA-mediated 
            silencing.
  JOURNAL   Nature. 2017 Feb 9;542(7640):197-202. doi: 10.1038/nature21025. Epub
            2017 Jan 23.
  PUBMED    28114302
REFERENCE   2  (bases 1 to 8763)
  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: 
            "10.1038/nature21025"; journalName: "Nature"; date: "2017-02-9- 9"; 
            volume: "542"; issue: "7640"; pages: "197-202"
COMMENT     Sequence Label: pcDNA3.1+_V5-ANKRD52
FEATURES             Location/Qualifiers
     source          1..8763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             907..948
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3683..3694
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4354..4371)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4360..4584
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4630..5058
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4717..4736)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4927..4948
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5067..5087)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5072..5401
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5252..5387
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5314..5333
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5468..6262
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5522..5541)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6132..6151
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6436..6557
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6473..6492)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6527..6546
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6606..6622)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6606..6622)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6619..6641)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6630..6646
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6654..6660,6661..6678,6679..6684))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6699..6720
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6837..6854)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7008..7596)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7088..7107)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7767..8558,8559..8627))
                     /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     8390..8409
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8628..8732)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 accatgggta agcctatccc taaccctctc ctcggtctcg attctacgat ggggatcctc
      961 agcatcacgg accagccgcc cctggtccag gccatcttta gccgagatgt ggaggaagtg
     1021 cgttccctac tctcgcagaa ggagaacatc aatgtgctgg accaagagag gcgaactcca
     1081 ttgcatgctg ctgcctacgt aggcgatgtc cccatcctcc agttgctact gatgtcaggt
     1141 gctaatgtca atgctaagga cacactgtgg ctgacccctc ttcatcgtgc tgctgcctcc
     1201 cgaaacgaga aggtgctggg gctgctgctg gcacattcag cagatgtgaa tgcccgggac
     1261 aagctgtggc agacaccact gcatgtggct gctgccaacc gggccaccaa gtgtgctgag
     1321 gctctggcac ccctgttgag cagcctcaac gtggctgaca ggagcgggcg cagtgccctg
     1381 caccatgcag tgcatagtgg gcatcttgag acggtgaacc tgctcctcaa caagggagcc
     1441 agcctgaatg tctgtgacaa aaaggagcgg cagcctctgc attgggcagc ttttctaggg
     1501 cacttggagg tcctaaaact gctggtggca cggggagcag acctcggctg caaggaccgc
     1561 aagggctatg ggctgctcca tacagctgct gccagtggcc agattgaagt ggtgaagtac
     1621 ctgcttcgga tgggagcgga gatcgatgaa cccaatgctt ttggaaacac agctttgcac
     1681 atcgcctgct acctgggcca ggatgctgtg gctattgagc tggtgaatgc cggagccaat
     1741 gtcaaccagc cgaatgacaa gggcttcacg ccactgcatg tggctgcagt ctcgaccaat
     1801 ggcgctctct gcttggagct actggttaat aatggggctg acgtcaacta ccagagcaaa
     1861 gaagggaaaa gtcctctgca catggctgca atccatggcc gtttcacacg ctcccagatc
     1921 ctcatccaga atggcagcga gattgattgt gccgacaaat ttgggaacac gccactgcat
     1981 gtggctgctc gatatggaca cgagctgctc atcagcaccc tcatgaccaa tggcgcagat
     2041 accgcccggc gtggcatcca tgacatgttc cccctgcact tagctgttct ctttggattc
     2101 tctgactgtt gtcgtaagct tctttcctca ggtcagttgt acagcattgt gtcttcactc
     2161 agcaatgagc atgtgctttc agctgggttt gacatcaata cacctgacaa ccttggccgt
     2221 acctgtcttc atgctgctgc ttccggaggg aatgttgaat gtcttaattt gctgttgagc
     2281 agtggagctg acttgaggag gagggacaaa tttggcagga ccccactgca ctatgcagct
     2341 gctaacggta gctaccagtg tgcagtaaca ttggtgactg ctggggcagg tgtcaacgag
     2401 gccgactgta aaggctgctc tcccctccac tacgctgccg cttctgacac ttacaggaga
     2461 gcggaacccc atacaccttc cagccatgat gccgaagagg acgagccact gaaggagtcc
     2521 cgcaggaagg aggccttctt ctgtctggag ttcttactgg ataacggtgc agacccctcc
     2581 ctgcgggaca ggcagggcta cacagctgtg cactatgcag ccgcctatgg caacagacag
     2641 aacctcgaac tgctcttaga aatgtccttt aactgcctgg aggatgtgga gagcaccatt
     2701 ccagtcagcc ctttgcactt agctgcctac aacggtcact gtgaagcctt gaagacgctg
     2761 gcggagacgc tggtgaatct ggacgtaagg gaccacaagg gccggaccgc actcttcctg
     2821 gccacggagc gcggctctac tgagtgtgtg gaggtgctta cagcccacgg cgcctctgcc
     2881 ctcatcaagg agcgcaagcg caagtggaca cccctgcacg ctgctgctgc ctctggccac
     2941 actgactccc tgcacttgct gatcgacagt ggggaacgag ctgacatcac agatgtcatg
     3001 gatgcctatg gacagacccc actgatgctg gccatcatga atggccatgt ggactgtgta
     3061 catctgctgc tagagaaagg atccacagct gatgctgctg acctccgggg ccgcactgcc
     3121 ctccaccgcg gggcagtgac tggctgtgag gactgcctgg ctgccctgct ggaccacgac
     3181 gcatttgtgc tgtgccgaga ctttaagggc cgcacgccca ttcacctggc ctcagcctgt
     3241 ggccacactg cagtactgcg gaccctgctg caggctgccc tttccacaga tcccctggat
     3301 gccggggtgg attacagcgg atactcgccc atgcactggg cctcctacac tggacatgaa
     3361 gattgtctgg agttgttact tgaacacagc ccgttttcgt acctggaagg aaaccccttc
     3421 actcctttgc actgtgcagt gattaataac caagacagca ccacagagat gctactggga
     3481 gctctgggtg ccaagattgt gaacagccga gatgccaaag gacggacccc ccttcacgcc
     3541 gctgccttcg cggacaatgt ctctgggctc cggatgctgc tgcagcatca agctgaggtg
     3601 aacgccactg accacactgg ccgcactgcg ctcatgacgg cggctgagaa cgggcagacc
     3661 gctgctgtgg aatttctgct gtatcgaggg aaggcagacc ttactgtgtt ggatgagaac
     3721 aagaacacgg ccctccactt ggcttgtagc aagggccatg agaaatgtgc cctcatgatc
     3781 ctggcagaaa cccaagacct tggccttatc aatgctacca acagtgcgct gcagatgcca
     3841 ctccacattg ctgcccggaa tggtctagct tctgtggtac aggccctgct gagtcatggg
     3901 gccacagtgc tggctgtgga tgaagaaggt cacaccccag cactggcctg tgcccccaac
     3961 aaagatgtgg cagactgcct ggccttgatc ctttccacca tgaagccttt cccacccaag
     4021 gacgccgtca gtcctttcag cttcagcctg ctcaagaact gcagcattgc agccgccaag
     4081 acggtgggtg gctgcggcgc cctgccccat ggggcctcct gcccctacag ccaggagcgg
     4141 cccggcgcca ttgggttaga tggctgctac tctgagtagc cccctccagt gtccctcccc
     4201 cgccggtggc ttgatatcta attctattta tttagaaaaa gtctaaacat ttagggcact
     4261 ttaaaggaga acacgactgg gtggaattct gcagatatcc agcacagtgg cggccgctcg
     4321 agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     4381 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     4441 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     4501 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     4561 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     4621 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4681 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4741 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4801 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     4861 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4921 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     4981 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     5041 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     5101 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     5161 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     5221 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     5281 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     5341 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     5401 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     5461 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     5521 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     5581 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     5641 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     5701 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     5761 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     5821 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     5881 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     5941 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     6001 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     6061 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     6121 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     6181 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     6241 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     6301 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     6361 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     6421 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     6481 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     6541 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     6601 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     6661 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     6721 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     6781 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6841 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6901 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6961 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     7021 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     7081 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     7141 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     7201 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     7261 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     7321 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     7381 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     7441 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7501 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7561 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7621 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7681 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7741 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7801 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7861 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7921 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7981 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     8041 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     8101 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     8161 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     8221 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     8281 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     8341 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     8401 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     8461 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     8521 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     8581 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8641 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     8701 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     8761 gtc
//
