LOCUS       sequence-481707-        9048 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 9048)
  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 9048)
  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 2
FEATURES             Location/Qualifiers
     source          1..9048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(37..57)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     534..553
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(563..892)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      577..712
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     877..897
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      985..1440
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1072..1091)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1282..1303
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1507..1526)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1626..1648
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1686..1704)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1772..1876
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1877..1945,1946..2737)
                     /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(2095..2114)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2908..3496
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3397..3416
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3573..3952
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          4248..5264
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     5188..5210
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     5272..5291
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             5382..5447
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    5466..5490
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             8565..8654
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     protein_bind    complement(8664..8688)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    8713..8834
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8724..8743)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8778..8797
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(9002..9048,1..553))
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
ORIGIN
        1 gcacctcgac gtcggcggtg acggtgaagc cgagccgctc gtagaagggg aggttgcggg
       61 gcgcggaggt ctccaggaag gcgggcaccc cggcgcgctc ggccgcctcc actccgggga
      121 gcacgacggc gctgcccaga cccttgccct ggtggtcggg cgagacgccg acggtggcca
      181 ggaaccacgc gggctccttg ggccggtgcg gcgccaggag gccttccatc tgttgctgcg
      241 cggccagccg ggaaccgctc aactcggcca tgcgcgggcc gatctcggcg aacaccgccc
      301 ccgcttcgac gctctccggc gtggtccaga ccgccaccgc ggcgccgtcg tccgcgaccc
      361 acaccttgcc gatgtcgagc ccgacgcgcg tgaggaagag ttcttgcagc tcggtgaccc
      421 gctcgatgtg gcggtccgga tcgacggtgt ggcgcgtggc ggggtagtcg gcgaacgcgg
      481 cggcgagggt gcgtacggcc ctggggacgt cgtcgcgggt ggcgaggcgc accgtgggct
      541 tgtactcggt catggtaagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact
      601 acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta
      661 gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag
      721 ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
      781 ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca
      841 tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac acattccaca
      901 ggtcgagatc cgggctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
      961 ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     1021 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     1081 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     1141 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     1201 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     1261 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     1321 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     1381 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     1441 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac
     1501 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     1561 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     1621 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     1681 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     1741 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     1801 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     1861 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     1921 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     1981 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     2041 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     2101 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     2161 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     2221 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     2281 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     2341 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     2401 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     2461 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     2521 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     2581 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     2641 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     2701 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     2761 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     2821 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2881 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2941 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3001 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3061 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3121 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3181 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3241 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3301 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3361 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3421 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3481 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3541 gccttttgct cacatggctc gacagatccc tcgacattga ttattgacta gttattaata
     3601 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     3661 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     3721 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     3781 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     3841 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     3901 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt
     3961 gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta
     4021 tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg gggggccgcc
     4081 catggggggg gcgggggggg gggggggggg gggggggggg gggggggggg ggggcgaggc
     4141 ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga
     4201 ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc
     4261 gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac
     4321 tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt
     4381 agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc
     4441 tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg
     4501 tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg
     4561 cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc
     4621 ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt
     4681 gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc cctccccgag
     4741 ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg
     4801 ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg
     4861 ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg
     4921 cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag ggacttcctt
     4981 tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc
     5041 gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc cttcgtgcgt
     5101 cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg ggggacggct
     5161 gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta
     5221 gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc
     5281 tggttattgt gctgtctcat cattttggca aagaattgat ttatcgattt gggaattcga
     5341 gctcggtacc agatcttcta gattcgaact cgagcaccat ggactacaaa gaccatgacg
     5401 gtgattataa agatcatgac atcgattaca aggatgacga tgacaagctc tgcagggcgg
     5461 ccatcacaag tttgtacaaa aaagcaggct ctttaaagga accaattcag tcgactggat
     5521 ccatgcccaa gggtgggtgc cctaaagcac cacagcagga agagcttccc ctcagcagcg
     5581 acatggtgga gaagcagact gggaaaaagg ataaagataa agtttctcta accaagaccc
     5641 caaaactgga gcgtggcgat ggcgggaagg aggtgaggga gcgagccagc aagcggaagc
     5701 tgcccttcac cgcgggcgcc aatggggagc agaaggactc ggacacagag aagcagggcc
     5761 ctgagcggaa gaggattaag aaggagcctg tcacccggaa ggccgggctg ctgtttggca
     5821 tggggctgtc tggaatccga gccggctacc ccctctccga gcgccagcag gtggcccttc
     5881 tcatgcagat gacggccgag gagtctgcca acagcccagt ggacacaaca ccaaagcacc
     5941 cctcccagtc tacagtgtgt cagaagggaa cgcccaactc tgcctcaaaa accaaagata
     6001 aagtgaacaa gagaaacgag cgtggagaga cccgcctgca ccgagccgcc atccgcgggg
     6061 acgcccggcg catcaaagag ctcatcagcg agggggcaga cgtcaacgtc aaggacttcg
     6121 caggctggac ggcgctgcac gaggcctgta accggggcta ctacgacgtc gcgaagcagc
     6181 tgctggctgc aggtgcggag gtgaacacca agggcctaga tgacgacacg cctttgcacg
     6241 acgctgccaa caacgggcac tacaaggtgg tgaagctgct gctgcggtac ggagggaacc
     6301 cgcagcagag caacaggaaa ggcgagacgc cgctgaaagt ggccaactcc cccacgatgg
     6361 tgaacctcct gttaggcaaa ggcacttaca cttccagcga ggagagctcg acggagagct
     6421 cagaagagga agacgcacca tccttcgcac cttccagttc agtcgacggc aacaacacgg
     6481 actccgagtt cgaaaaaggc ctcaagcaca aggccaagaa cccagagcca cagaaggcca
     6541 cggcccccgt caaggacgag tatgagtttg atgaggacga cgagcaggac agggttcctc
     6601 cggtggacga caagcaccta ttgaaaaagg actacagaaa agaaacgaaa tccaatagtt
     6661 ttatctctat acccaaaatg gaggttaaaa gttacactaa aaataacacg attgcaccaa
     6721 agaaagcgtc ccatcgtatc ctgtcagaca cgtcggacga ggaggacgcg agtgtcaccg
     6781 tggggacagg agagaagctg agactctcgg cacatacgat attgcctggt agtaagacac
     6841 gagagccttc taatgccaag cagcagaagg aaaaaaataa agtgaaaaag aagcgaaaga
     6901 aagaaacaaa aggcagagag gttcgcttcg gaaagcggag cgacaagttc tgctcctcgg
     6961 agtcggagag cgagtcctca gagagtgggg aggatgacag ggactctctg gggagctctg
     7021 gctgcctcaa ggggtccccg ctggtgctga aggacccctc cctgttcagc tccctctctg
     7081 cctcctccac ctcgtctcac gggagctctg ccgcccagaa gcagaacccc agccacacag
     7141 accagcacac caagcactgg cggacagaca attggaaaac catttcttcc ccggcttggt
     7201 cagaggtcag ttctttatca gactccacaa ggacgagact gacaagcgag tctgactact
     7261 cctctgaggg ctccagtgtg gaatcgctga agccagtgag gaagaggcag gagcacagga
     7321 agcgagcctc cctgtcggag aagaagagcc ccttcctgtc cagcgcggag ggcgctgtcc
     7381 ccaaactgga caaggagggg aaagttgtca aaaaacataa aacaaaacac aaacacaaaa
     7441 acaaggagaa gggacagtgt tccatcagcc aagagctgaa gttgaaaagt tttacttacg
     7501 aatatgagga ctccaagcag aagtcagata aggctatact gttagagaat gatctttcca
     7561 ctgaaaacaa gctaaaagtg ttaaagcacg atcgcgacca ctttaaaaaa gaagagaaac
     7621 ttagcaaaat gaaattagaa gaaaaagaat ggctctttaa agatgaaaaa tcactgaaga
     7681 gaatcaaaga cacaaacaaa gacatcagca ggtctttccg agaagagaaa gaccgttcga
     7741 ataaagcaga aaaggagaga tcgctgaagg aaaagtctcc gaaagaagaa aaactgagac
     7801 tgtacaaaga ggagagaaag aagaaatcaa aagaccggcc ctcaaaatta gagaagaaga
     7861 atgatttaaa agaggacaaa atttcaaaag agaaggagaa gatttttaaa gaagataaag
     7921 aaaaactcaa aaaagaaaag gtttataggg aagattctgc ttttgacgaa tattgtaaca
     7981 aaaatcagtt tctggagaat gaagacacca aatttagcct ttctgacgat cagcgagatc
     8041 ggtggttttc tgacttgtcc gattcatcct ttgatttcaa aggggaggac agctgggact
     8101 cgccagtgac agactacagg gacatgaaga gcgactctgt ggccaagctc atcttggaga
     8161 cggtgaagga ggacagcaag gagaggaggc gggacagccg ggcccgggag aagcgagact
     8221 acagagagcc cttcttccga aagaaggaca gggactattt ggataaaaac tctgagaaga
     8281 ggaaagagca gactgaaaag cataaaagtg tccctggcta cctttcggaa aaggacaaga
     8341 agaggagaga gtccgcagag gccgggcggg acagaaagga cgccctggag agctgcaagg
     8401 agcgcaggga cggcagggcc aagcccgagg aggtgcaccg ggaggagctg aaggagtgtg
     8461 gctgcgagag tggcttcaag gacaagtccg acggcgactt tgggaagggc ctggagccgt
     8521 gggaacggca ccacccagca gcggccgcac tcgagatggc cgcataccca tacgatgttc
     8581 cagattacgc tggctatccc tatgacgtcc cggactatgc aggatcctat ccatatgacg
     8641 ttccagatta cgctgatatc tagacccagc tttcttgtac aaagtggtga tggccgcttc
     8701 gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     8761 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     8821 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga
     8881 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc
     8941 tcgacgatgc atggggtcgt gcgctccttt cggtcgggcg ctgcgggtcg tggggcgggc
     9001 gtcaggcacc gggcttgcgg gtcatgcacc aggtgcgcgg tccttcgg
//
