LOCUS       sequence-474270-        7454 bp    DNA     circular SYN 02-JUN-2025
DEFINITION  Expresses human WDR5 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7454)
  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, 112699
REFERENCE   2  (bases 1 to 7454)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "iScience 
            2025 May 19, 112699"
COMMENT     Sequence Label: pCAG-puro-9xmyc-WDR5
FEATURES             Location/Qualifiers
     source          1..7454
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      669..1257
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1158..1177
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1334..1713
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     gap             2008..2017
                     /estimated_length=10
     intron          2243..3259
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3183..3205
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3267..3286
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3377..3406
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3413..3442
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3449..3478
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3497..3526
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3533..3562
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3569..3598
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3617..3646
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3653..3682
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3689..3718
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    3752..3776
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4831..4855)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    4880..5001
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4891..4910)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4945..4964
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5169..5768)
                     /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(5252..5272)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     5749..5768
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(5778..6107)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5792..5927
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6092..6112
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6200..6655
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6287..6306)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6497..6518
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6722..6741)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6841..6863
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6901..6919)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6987..7091
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7092..7160,7161..7454,1..498)
                     /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(7310..7329)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
       61 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
      121 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
      181 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
      241 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
      301 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
      361 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
      421 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
      481 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
      541 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
      601 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
      661 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
      721 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
      781 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
      841 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
      901 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
      961 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     1021 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     1081 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     1141 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     1201 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     1261 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatggct
     1321 cgacagatcc ctcgacattg attattgact agttattaat agtaatcaat tacggggtca
     1381 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     1441 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     1501 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     1561 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     1621 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     1681 tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc
     1741 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta
     1801 ttttgtgcag cgatgggggc gggggggggg ggggggcgcg ccccaggggg gggggggggg
     1861 gggggggggg gggggggggg gggggggggg gggggggggg cggcccagtg ggggcggggg
     1921 gggggggggg gttctttttt tgcggggcgg ggggggcggg ggccccccaa aaaacagcaa
     1981 ccggcggggg gggggggggg gggggcgnnn nnnnnnncga cccctatttt gttttttttt
     2041 tttttttttt aattgtgggg gggggggggg gggggggggg ggggggggcg cgcggggggg
     2101 gggggggggg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca
     2161 gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa
     2221 aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc
     2281 cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag
     2341 cgggcgggac ggcccttctc ctccgggctg taattagcgc ttggtttaat gacggcttgt
     2401 ttcttttctg tggctgcgtg aaagccttga ggggctccgg gagggccctt tgtgcggggg
     2461 gagcggctcg gggggtgcgt gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc
     2521 gctgcccggc ggctgtgagc gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt
     2581 gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa
     2641 caaaggctgc gtgcggggtg tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt
     2701 cgggctgcaa ccccccctgc acccccctcc ccgagttgct gagcacggcc cggcttcggg
     2761 tgcggggctc cgtacggggc gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca
     2821 ggtgggggtg ccgggcgggg cggggccgcc tcgggccggg gagggctcgg gggaggggcg
     2881 cggcggcccc cggagcgccg gcggctgtcg aggcgcggcg agccgcagcc attgcctttt
     2941 atggtaatcg tgcgagaggg cgcagggact tcctttgtcc caaatctgtg cggagccgaa
     3001 atctgggagg cgccgccgca ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg
     3061 caggaaggaa atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc cccttctccc
     3121 tctccagcct cggggctgtc cgcgggggga cggctgcctt cgggggggac ggggcagggc
     3181 ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc atgttcatgc
     3241 cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt
     3301 tggcaaagaa ttgatttatc gatttgggaa ttcgagctcg gtaccagatc ttctagattc
     3361 gaactcgagc accatggaac aaaagttgat ttctgaagaa gatttgaacg gtgaacaaaa
     3421 gctaatctcc gaggaagact tgaacggtga acaaaaatta atctcagaag aagacttgaa
     3481 cggatcctct agaggtgaac aaaagttgat ttctgaagaa gatttgaacg gtgaacaaaa
     3541 gctaatctcc gaggaagact tgaacggtga acaaaaatta atctcagaag aagacttgaa
     3601 cggatcctct agaggtgaac aaaagttgat ttctgaagaa gatttgaacg gtgaacaaaa
     3661 gctaatctcc gaggaagact tgaacggtga acaaaaatta atctcagaag aagacttgaa
     3721 cggatccact agcctctgca gggcggccat cacaagtttg tacaaaaaag caggctcttt
     3781 aaaggaacca attcagtcga ctatggcgac ggaggagaag aagcccgaga ccgaggccgc
     3841 cagagcacag ccaacccctt cgtcatccgc cactcagagc aagcctacac ctgtgaagcc
     3901 aaactatgct ctaaagttca cccttgctgg ccacaccaaa gcagtgtcct ccgtgaaatt
     3961 cagcccgaat ggagagtggc tggcaagttc atctgctgat aaacttatta aaatttgggg
     4021 cgcgtatgat gggaaatttg agaaaaccat atctggtcac aagctgggaa tatccgatgt
     4081 agcctggtcg tcagattcta accttcttgt ttctgcctca gatgacaaaa ccttgaagat
     4141 atgggacgtg agctcgggca agtgtctgaa aaccctgaag ggacacagta attatgtctt
     4201 ttgctgcaac ttcaatcccc agtccaacct tattgtctca ggatcctttg acgaaagcgt
     4261 gaggatatgg gatgtgaaaa cagggaagtg cctcaagact ttgccagctc actcggatcc
     4321 agtctcggcc gttcatttta atcgtgatgg atccttgata gtttcaagta gctatgatgg
     4381 tctctgtcgc atctgggaca ccgcctcagg ccagtgcctg aagacgctca tcgatgacga
     4441 caaccccccc gtgtcttttg tgaagttctc cccgaacggc aaatacatcc tggccgccac
     4501 gctggacaac actctgaagc tctgggacta cagcaagggg aagtgcctga agacgtacac
     4561 tggccacaag aatgagaaat actgcatatt tgccaatttc tctgttactg gtgggaagtg
     4621 gattgtgtct ggctcagagg ataaccttgt ttacatctgg aaccttcaga cgaaagagat
     4681 tgtacagaaa ctacaaggcc acacagatgt cgtgatctca acagcttgtc acccaacaga
     4741 aaacatcatc gcctctgctg cgctagaaaa tgacaaaaca attaaactgt ggaagagtga
     4801 ctgctaagcg gccgcactcg agatatctag acccagcttt cttgtacaaa gtggtgatgg
     4861 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     4921 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     4981 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     5041 ggggagatgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat
     5101 aaggatctcg acgatgcatg gggtcgtgcg ctcctttcgg tcgggcgctg cgggtcgtgg
     5161 ggcgggcgtc aggcaccggg cttgcgggtc atgcaccagg tgcgcggtcc ttcgggcacc
     5221 tcgacgtcgg cggtgacggt gaagccgagc cgctcgtaga aggggaggtt gcggggcgcg
     5281 gaggtctcca ggaaggcggg caccccggcg cgctcggccg cctccactcc ggggagcacg
     5341 acggcgctgc ccagaccctt gccctggtgg tcgggcgaga cgccgacggt ggccaggaac
     5401 cacgcgggct ccttgggccg gtgcggcgcc aggaggcctt ccatctgttg ctgcgcggcc
     5461 agccgggaac cgctcaactc ggccatgcgc gggccgatct cggcgaacac cgcccccgct
     5521 tcgacgctct ccggcgtggt ccagaccgcc accgcggcgc cgtcgtccgc gacccacacc
     5581 ttgccgatgt cgagcccgac gcgcgtgagg aagagttctt gcagctcggt gacccgctcg
     5641 atgtggcggt ccggatcgac ggtgtggcgc gtggcggggt agtcggcgaa cgcggcggcg
     5701 agggtgcgta cggccctggg gacgtcgtcg cgggtggcga ggcgcaccgt gggcttgtac
     5761 tcggtcatgg taagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc
     5821 tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag
     5881 ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg
     5941 ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
     6001 gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt
     6061 ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt ccacaggtcg
     6121 agatccgggc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     6181 cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     6241 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     6301 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     6361 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     6421 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     6481 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     6541 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     6601 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttcctga
     6661 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
     6721 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     6781 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     6841 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     6901 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     6961 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     7021 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     7081 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     7141 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     7201 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     7261 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     7321 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     7381 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     7441 taagagaatt atgc
//
