LOCUS       sequence-481703-        8474 bp    DNA     circular SYN 23-SEP-2025
DEFINITION  Expresses human ANKRD11 fragment in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8474)
  AUTHORS   Sashiyama S, Nakagawa T, Nakagawa M, Hosogane M, Watanabe Y, 
            Ashitomi H, Yamane K, Shibuya N, Moroishi T, Nakayama K, Hosoi T
  TITLE     KBG syndrome-associated protein ANKRD11 regulates SETD5 expression 
            to modulate rRNA levels and translation.
  JOURNAL   iScience. 2025 May 19;28(6):112699. doi: 10.1016/j.isci.2025.112699.
            eCollection 2025 Jun 20.
  PUBMED    40520101
REFERENCE   2  (bases 1 to 8474)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 23, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "iScience.";
            date: "2025-05-19"; volume: "28(6):112699. doi"; pages: " 
            10.1016/j.isci.2025.112699. eCollection 2025 Jun 20"
COMMENT     Sequence Label: pCAG-puro-3xFLAG-ANKRD11-3xHA fragment 6
FEATURES             Location/Qualifiers
     source          1..8474
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          293..1309
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1233..1255
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1317..1336
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1427..1492
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    1511..1535
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4079..4168
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     protein_bind    complement(4194..4218)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    4243..4364
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4254..4273)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4308..4327
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4532..5131)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4615..4635)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     5112..5131
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(5141..5470)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5155..5290
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5455..5475
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5563..6018
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5650..5669)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5860..5881
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6085..6104)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6204..6226
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6264..6282)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6350..6454
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6455..6523,6524..7315)
                     /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(6673..6692)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7486..8074
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7975..7994
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        join(8151..8474,1..56)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg
       61 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt
      121 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcgaggggc ggggcggggc
      181 gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat
      241 ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc
      301 tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct
      361 ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg
      421 taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg aaagccttga
      481 ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt gcgtgtgtgt
      541 gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc gctgcgggcg
      601 cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg ggggcggtgc
      661 cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg tgtgcgtggg
      721 ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc acccccctcc
      781 ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc gtggcgcggg
      841 gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg cggggccgcc
      901 tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg gcggctgtcg
      961 aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg cgcagggact
     1021 tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca ccccctctag
     1081 cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg agggccttcg
     1141 tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc cgcgggggga
     1201 cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg
     1261 ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc tcctgggcaa
     1321 cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttgatttatc gatttgggaa
     1381 ttcgagctcg gtaccagatc ttctagattc gaactcgagc accatggact acaaagacca
     1441 tgacggtgat tataaagatc atgacatcga ttacaaggat gacgatgaca agctctgcag
     1501 ggcggccatc acaagtttgt acaaaaaagc aggctcttta aaggaaccaa ttcagtcgac
     1561 tggatccggt accgaattcg acagggcgcc cctgcccccg gttcccgcgg agaagtttgc
     1621 ctgcttgtcg ccagggtact actccccaga ctatggcctc ccgtcgccca aagtcgacgc
     1681 tttgcactgc ccaccggctg ccgttgtcac tgtcaccccg tctccagagg gcgtcttctc
     1741 aagtttacaa gcaaaacctt ccccttcccc cagagccgag ctgctggttc cttccctcga
     1801 aggggccctt cccccggacc tggacacctc cgaggaccag caggcgacgg ccgccatcat
     1861 ccccccggag cccagctacc tggagccgct ggacgagggt cccttcagcg ccgtcatcac
     1921 cgaggagccc gttgagtggg cccacccctc cgagcaggcg cttgcctcta gcctgatcgg
     1981 gggcacctct gaaaaccctg tgagctggcc tgtgggctcg gacctcctgc tgaagtctcc
     2041 acagagattc cccgagtccc caaagcgttt ctgccccgcg gaccccctcc actctgccgc
     2101 cccagggccc ttcagcgcct cggaggcgcc gtaccccgcc cctcccgcct ctcctgcccc
     2161 gtacgctctg cccgtcgctg agccggggct ggaggacgtc aaggacggag tggacgccgt
     2221 ccccgccgcc atctccacct cagaggcggc tccctacgcc cctccctccg ggctggagtc
     2281 cttcttcagc aactgcaagt cacttccgga agccccgctg gacgtggccc ccgagcccgc
     2341 ctgtgtagcc gctgtggctc aggtggaggc tctggggccc ctggaaaata gcttcctgga
     2401 cggcagccgc ggcctgtctc acctcggcca ggtggagccg gtgccctggg cggacgcctt
     2461 cgccggcccc gaggacgacc tggacctggg gcccttctcc ctgccggagc ttcccctgca
     2521 gactaaagat gccgcagatg gtgaagcgga acccgtggaa gaaagtcttg ctcctccaga
     2581 agagatgcct ccaggggccc ccggggtcat aaacggtggg gatgtttcca ccgtagtggc
     2641 tgaggagccg ccggcactgc ctcctgacca ggcctccacc cggctccctg cagagctcga
     2701 gcctgagccc tcaggggagc caaagctgga cgtggctcta gaagctgcgg tggaggcgga
     2761 gacggtgccg gaagagaggg cccgtgggga tccggactcc agcgtggagc ccgcgcccgt
     2821 tcccccagaa cagcgcccac tggggagcgg agaccagggg gctgaggctg aaggcccccc
     2881 cgccgcgtcc ctctgtgccc ctgacggccc cgccccgaac actgtggcac aagctcaggc
     2941 cgcagacggt gccggccccg aggacgacac tgaggcctcc cgtgccgccg ccccagccga
     3001 aggccctcct ggcggcatcc agccggaagc cgcagaacca aaacccacgg ccgaagcccc
     3061 gaaggccccc cgagtggagg agatccctca gcgcatgacc aggaaccggg cgcagatgct
     3121 cgcgaaccag agcaagcagg gcccgccccc ctccgagaag gagtgcgccc ccacccctgc
     3181 cccggtcacc agggccaagg cccgcggctc cgaggacgac gacgcccagg cccagcatcc
     3241 gcgcaaacgc cgctttcagc gctccaccca gcagctgcag cagcagctga acacgtccac
     3301 gcagcagacg cgggaggtga tccagcagac gctggccgcc atcgtggacg ccatcaagct
     3361 ggatgccatc gagccctacc acagcgacag ggccaacccc tacttcgaat acctgcagat
     3421 caggaagaag atcgaggaga agcgcaagat cctgtgctgt atcacgccgc aggcgcccca
     3481 gtgctacgcc gagtacgtca cctacacggg ctcctacctc ctggacggca agccgctcag
     3541 caagctccac atccccgtga tcgcaccccc tccctccctg gcggagcccc tgaaggagct
     3601 gttcaggcag caggaggccg tccggggaaa gctgcgtcta cagcacagca tcgagcggga
     3661 gaagctgatc gtatcctgtg agcaggagat tctgcgggtt cactgccggg cggccaggac
     3721 catcgccaac caggcagtgc cattcagcgc ctgcacgatg ctgctggact ccgaggtcta
     3781 caacatgccc ctggagagcc agggtgacga gaacaagtca gtgcgcgacc gtttcaacgc
     3841 ccgccagttc atctcctggc tccaggacgt ggatgacaag tatgaccgca tgaagacttg
     3901 cctcctcatg cggcagcagc acgaggccgc ggccctgaac gccgtgcaga ggatggagtg
     3961 gcagctgaag gtgcaggaac tggaccccgc cgggcacaag tccctgtgcg tgaacgaggt
     4021 gccctccttc tacgtgccca tggtcgacgt caacgacgac tttgtattgt tgccggcata
     4081 cccatacgat gttccagatt acgctggcta tccctatgac gtcccggact atgcaggatc
     4141 ctatccatat gacgttccag attacgcttg gcggccgcac tcgagatatc tagacccagc
     4201 tttcttgtac aaagtggtga tggccgcttc gagcagacat gataagatac attgatgagt
     4261 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     4321 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     4381 ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc
     4441 tctacaaatg tggtaaaatc gataaggatc tcgacgatgc atggggtcgt gcgctccttt
     4501 cggtcgggcg ctgcgggtcg tggggcgggc gtcaggcacc gggcttgcgg gtcatgcacc
     4561 aggtgcgcgg tccttcgggc acctcgacgt cggcggtgac ggtgaagccg agccgctcgt
     4621 agaaggggag gttgcggggc gcggaggtct ccaggaaggc gggcaccccg gcgcgctcgg
     4681 ccgcctccac tccggggagc acgacggcgc tgcccagacc cttgccctgg tggtcgggcg
     4741 agacgccgac ggtggccagg aaccacgcgg gctccttggg ccggtgcggc gccaggaggc
     4801 cttccatctg ttgctgcgcg gccagccggg aaccgctcaa ctcggccatg cgcgggccga
     4861 tctcggcgaa caccgccccc gcttcgacgc tctccggcgt ggtccagacc gccaccgcgg
     4921 cgccgtcgtc cgcgacccac accttgccga tgtcgagccc gacgcgcgtg aggaagagtt
     4981 cttgcagctc ggtgacccgc tcgatgtggc ggtccggatc gacggtgtgg cgcgtggcgg
     5041 ggtagtcggc gaacgcggcg gcgagggtgc gtacggccct ggggacgtcg tcgcgggtgg
     5101 cgaggcgcac cgtgggcttg tactcggtca tggtaagctt tttgcaaaag cctaggcctc
     5161 caaaaaagcc tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg
     5221 cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta
     5281 ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc
     5341 tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat
     5401 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta
     5461 actgacacac attccacagg tcgagatccg ggctggcgta atagcgaaga ggcccgcacc
     5521 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg
     5581 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     5641 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     5701 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     5761 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     5821 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     5881 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     5941 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     6001 tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     6061 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     6121 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     6181 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     6241 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     6301 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     6361 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     6421 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     6481 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     6541 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     6601 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     6661 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     6721 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     6781 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     6841 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     6901 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     6961 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     7021 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     7081 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     7141 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     7201 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     7261 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     7321 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     7381 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     7441 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     7501 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     7561 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     7621 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     7681 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     7741 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     7801 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     7861 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     7921 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     7981 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     8041 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     8101 gttcctggcc ttttgctggc cttttgctca catggctcga cagatccctc gacattgatt
     8161 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     8221 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     8281 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     8341 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     8401 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     8461 ccagtacatg acct
//
