LOCUS       sequence-481705-        8000 bp    DNA     circular SYN 18-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 8000)
  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 8000)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 18, 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 1
FEATURES             Location/Qualifiers
     source          1..8000
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     476..495
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(505..834)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      519..654
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     819..839
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      927..1382
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1014..1033)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1224..1245
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1449..1468)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1568..1590
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1628..1646)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1714..1818
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1819..1887,1888..2679)
                     /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(2037..2056)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2850..3438
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3339..3358
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3515..3894
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          4186..5202
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     5126..5148
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     5210..5229
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             5320..5385
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    5404..5428
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             7459..7548
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     protein_bind    complement(7558..7582)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    7607..7728
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7618..7637)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7672..7691
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(7896..8000,1..495))
                     /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(7979..7999)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 gggcgcggag gtctccagga aggcgggcac cccggcgcgc tcggccgcct ccactccggg
       61 gagcacgacg gcgctgccca gacccttgcc ctggtggtcg ggcgagacgc cgacggtggc
      121 caggaaccac gcgggctcct tgggccggtg cggcgccagg aggccttcca tctgttgctg
      181 cgcggccagc cgggaaccgc tcaactcggc catgcgcggg ccgatctcgg cgaacaccgc
      241 ccccgcttcg acgctctccg gcgtggtcca gaccgccacc gcggcgccgt cgtccgcgac
      301 ccacaccttg ccgatgtcga gcccgacgcg cgtgaggaag agttcttgca gctcggtgac
      361 ccgctcgatg tggcggtccg gatcgacggt gtggcgcgtg gcggggtagt cggcgaacgc
      421 ggcggcgagg gtgcgtacgg ccctggggac gtcgtcgcgg gtggcgaggc gcaccgtggg
      481 cttgtactcg gtcatggtaa gctttttgca aaagcctagg cctccaaaaa agcctcctca
      541 ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat
      601 tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg
      661 agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc
      721 tgctggggag cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg
      781 catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca
      841 caggtcgaga tccgggctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
      901 agttgcgcag cctgaatggc gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg
      961 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     1021 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     1081 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     1141 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     1201 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     1261 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     1321 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     1381 ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
     1441 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     1501 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     1561 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     1621 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     1681 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     1741 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     1801 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     1861 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     1921 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     1981 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     2041 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     2101 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     2161 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     2221 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     2281 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     2341 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     2401 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     2461 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     2521 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     2581 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     2641 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     2701 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     2761 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2821 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2881 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2941 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     3001 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3061 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3121 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3181 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3241 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3301 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3361 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3421 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3481 tggccttttg ctcacatggc tcgacagatc cctcgacatt gattattgac tagttattaa
     3541 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     3601 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     3661 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     3721 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     3781 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     3841 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag
     3901 gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg
     3961 tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg gggggggggg
     4021 gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg ggcgaggcgg
     4081 agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg
     4141 cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc
     4201 tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg
     4261 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag
     4321 cgcttggttt aatgacggct tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc
     4381 cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg
     4441 gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg
     4501 gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg
     4561 tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga
     4621 gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc tccccgagtt
     4681 gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc
     4741 gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc
     4801 ggggagggct cgggggaggg gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg
     4861 gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg
     4921 tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc
     4981 ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg
     5041 ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg ggacggctgc
     5101 cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga
     5161 gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg
     5221 gttattgtgc tgtctcatca ttttggcaaa gaattgattt atcgatttgg gaattcgagc
     5281 tcggtaccag atcttctaga ttcgaactcg agcaccatgg actacaaaga ccatgacggt
     5341 gattataaag atcatgacat cgattacaag gatgacgatg acaagctctg cagggcggcc
     5401 atcacaagtt tgtacaaaaa agcaggctct ttaaaggaac caattcagtc gactggatcc
     5461 atgcccaagg gtgggtgccc taaagcacca cagcaggaag agcttcccct cagcagcgac
     5521 atggtggaga agcagactgg gaaaaaggat aaagataaag tttctctaac caagacccca
     5581 aaactggagc gtggcgatgg cgggaaggag gtgagggagc gagccagcaa gcggaagctg
     5641 cccttcaccg cgggcgccaa tggggagcag aaggactcgg acacagagaa gcagggccct
     5701 gagcggaaga ggattaagaa ggagcctgtc acccggaagg ccgggctgct gtttggcatg
     5761 gggctgtctg gaatccgagc cggctacccc ctctccgagc gccagcaggt ggcccttctc
     5821 atgcagatga cggccgagga gtctgccaac agcccagtgg acacaacacc aaagcacccc
     5881 tcccagtcta cagtgtgtca gaagggaacg cccaactctg cctcaaaaac caaagataaa
     5941 gtgaacaaga gaaacgagcg tggagagacc cgcctgcacc gagccgccat ccgcggggac
     6001 gcccggcgca tcaaagagct catcagcgag ggggcagacg tcaacgtcaa ggacttcgca
     6061 ggctggacgg cgctgcacga ggcctgtaac cggggctact acgacgtcgc gaagcagctg
     6121 ctggctgcag gtgcggaggt gaacaccaag ggcctagatg acgacacgcc tttgcacgac
     6181 gctgccaaca acgggcacta caaggtggtg aagctgctgc tgcggtacgg agggaacccg
     6241 cagcagagca acaggaaagg cgagacgccg ctgaaagtgg ccaactcccc cacgatggtg
     6301 aacctcctgt taggcaaagg cacttacact tccagcgagg agagctcgac ggagagctca
     6361 gaagaggaag acgcaccatc cttcgcacct tccagttcag tcgacggcaa caacacggac
     6421 tccgagttcg aaaaaggcct caagcacaag gccaagaacc cagagccaca gaaggccacg
     6481 gcccccgtca aggacgagta tgagtttgat gaggacgacg agcaggacag ggttcctccg
     6541 gtggacgaca agcacctatt gaaaaaggac tacagaaaag aaacgaaatc caatagtttt
     6601 atctctatac ccaaaatgga ggttaaaagt tacactaaaa ataacacgat tgcaccaaag
     6661 aaagcgtccc atcgtatcct gtcagacacg tcggacgagg aggacgcgag tgtcaccgtg
     6721 gggacaggag agaagctgag actctcggca catacgatat tgcctggtag taagacacga
     6781 gagccttcta atgccaagca gcagaaggaa aaaaataaag tgaaaaagaa gcgaaagaaa
     6841 gaaacaaaag gcagagaggt tcgcttcgga aagcggagcg acaagttctg ctcctcggag
     6901 tcggagagcg agtcctcaga gagtggggag gatgacaggg actctctggg gagctctggc
     6961 tgcctcaagg ggtccccgct ggtgctgaag gacccctccc tgttcagctc cctctctgcc
     7021 tcctccacct cgtctcacgg gagctctgcc gcccagaagc agaaccccag ccacacagac
     7081 cagcacacca agcactggcg gacagacaat tggaaaacca tttcttcccc ggcttggtca
     7141 gaggtcagtt ctttatcaga ctccacaagg acgagactga caagcgagtc tgactactcc
     7201 tctgagggct ccagtgtgga atcgctgaag ccagtgagga agaggcagga gcacaggaag
     7261 cgagcctccc tgtcggagaa gaagagcccc ttcctgtcca gcgcggaggg cgctgtcccc
     7321 aaactggaca aggaggggaa agttgtcaaa aaacataaaa caaaacacaa acacaaaaac
     7381 aaggagaagg gacagtgttc catcagccaa gagctgaagt tgaaaagttt tactgcggcc
     7441 gcactcgaga tggccgcata cccatacgat gttccagatt acgctggcta tccctatgac
     7501 gtcccggact atgcaggatc ctatccatat gacgttccag attacgctga tatctagacc
     7561 cagctttctt gtacaaagtg gtgatggccg cttcgagcag acatgataag atacattgat
     7621 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     7681 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     7741 tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta aagcaagtaa
     7801 aacctctaca aatgtggtaa aatcgataag gatctcgacg atgcatgggg tcgtgcgctc
     7861 ctttcggtcg ggcgctgcgg gtcgtggggc gggcgtcagg caccgggctt gcgggtcatg
     7921 caccaggtgc gcggtccttc gggcacctcg acgtcggcgg tgacggtgaa gccgagccgc
     7981 tcgtagaagg ggaggttgcg
//
