LOCUS       sequence-235214-        8178 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a Flag-tagged version of DDX11 KAE mutant in mammalian
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8178)
  AUTHORS   Cortone G, Zheng G, Pensieri P, Chiappetta V, Tate R, Malacaria E, 
            Pichierri P, Yu H, Pisani FM
  TITLE     Interaction of the Warsaw breakage syndrome DNA helicase DDX11 with 
            the replication fork-protection factor Timeless promotes sister 
            chromatid cohesion.
  JOURNAL   PLoS Genet. 2018 Oct 10;14(10):e1007622. doi: 
            10.1371/journal.pgen.1007622. eCollection 2018 Oct.
  PUBMED    30303954
REFERENCE   2  (bases 1 to 8178)
  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"; journalName: "PLoS 
            Genet."; date: "2018-10-10"; volume: "14(10):e1007622. doi"; pages: 
            " 10.1371/journal.pgen.1007622. eCollection 2018 Oct"
COMMENT     Sequence Label: pcDNA-DDX11-Flag-KAE
FEATURES             Location/Qualifiers
     source          1..8178
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1567..1578
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     CDS             3637..3702
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(3733..3751)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3734..3751)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3771..3788)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3777..4001
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4047..4475
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4134..4153)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4344..4365
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4484..4504)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4489..4818
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4669..4804
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4731..4750
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4885..5679
                     /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(4939..4958)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5549..5568
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5853..5974
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5890..5909)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5944..5963
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6023..6039)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6023..6039)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6036..6058)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6047..6063
                     /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(6071..6077,6078..6095,6096..6101))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6116..6137
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6254..6271)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6425..7013)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6505..6524)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7184..7975,7976..8044))
                     /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     7807..7826
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8045..8149)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagcttatgc
      901 atcatcaccc atcaccacat ggctaatgaa acacagaagg ttggtgccat ccattttcct
      961 tttcccttca caccctattc catccaggaa gacttcatgg cagagctgta ccgggttttg
     1021 gaggctggca agattgggat atttgagagt ccaactggca ctgggaagtc cttaagtctt
     1081 atttgtgggg ccctctcttg gctccgtgac tttgaacaga agaagcgtga agaagaggca
     1141 cgactccttg aaactggaac tggcccctta catgatgaga aagatgaatc cctgtgtctg
     1201 tcttcttcct gcgaaggggc tgcaggcacc ccgaggcctg ctggagaacc ggcctgggtt
     1261 actcagtttg tgcagaagaa agaagagagg gacctggtgg accgactaaa ggcggagcag
     1321 gccaggagga agcagcgaga agaacgcctg cagcagctgc agcacagggt gcagctcaag
     1381 tatgcagcca agcgcctgag gcaggaagaa gaagaaagag agaatctcct ccgcctcagc
     1441 agggagatgc tagagacagg cccggaggct gagcggctgg agcagctgga gtctggggag
     1501 gaggagctgg tcctcgccaa agccgagagt gatgaggaga aaaaggtggc gagcagagtg
     1561 gatgaggatg aggatgacct ggaggaagaa cacataacta agatttatta ctgtagtcgg
     1621 acacactccc agctggccca gtttgtgcat gaggtgaaga agagcccctt tggcaaggat
     1681 gttcggctgg tctcccttgg ctcccggcag aacctttgtg taaatgaaga cgtgaaaagc
     1741 ctaggttctg tgcagcttat caacgaccgc tgtgtggaca tgcagagaag caggcacgag
     1801 aagaagaaag gagctgagga ggagaagcca aagaggagga ggcaggagaa gcaggcagcc
     1861 tgccccttct acaaccacga gcagatgggc cttctccggg atgaggccct ggcagaggtg
     1921 aaggacatgg agcagctgct ggcccttggg aaggaggccc gggcctgtcc ctattacggg
     1981 agccgccttg ccatccctgc agcccagctg gtggtgctgc cctatcagat gctgctgcat
     2041 gcggccactc ggcaggccgc gggcatccgg ctgcaggacc aggtggtgat catcgacgag
     2101 gcgcacaacc tgatcgacac catcacgggc atgcacagcg tggaggtcag cggctcccag
     2161 ctctgccagg cccattccca gctgctgcag tacgtggagc gatacgggaa gcgtttgaag
     2221 gccaagaacc tgatgtacct gaagcagatc ctgtatttgc tggagaaatt cgtggctgtg
     2281 ctagggggga acattaagca aaatcccaat acacagagtc tgtcacagac agggacggag
     2341 ctgaagacca tcaacgactt tctcttccag agccagatcg acaacatcaa cctgttcaag
     2401 gtgcagcgat actgtgagaa gagcatgatc agcagaaagc tctttggatt cactgaacgg
     2461 tacggagcag tgttctcatc ccgggagcag cccaaactgg ctgggtttca gcaattcctg
     2521 cagagcctgc agcccaggac gactgaagct cttgcagccc ctgcagacga gagtcaggcc
     2581 agcaccctgc gaccagcttc tccactgatg cacattgaag gcttcctggc agctctcact
     2641 acggccaacc aggacggcag ggtcatcctg agccgccaag gcagcctcag tcagagcacc
     2701 ctgaagtttt tgctcctgaa tccagctgtg cactttgccc aagtggtgaa ggaatgccgg
     2761 gcagtggtca ttgcgggggg taccatgcag ccggtgtcag acttccggca gcagctgctg
     2821 gcctgtgccg gggtggaagc tgagcgcgtg gtggagtttt cctgtggtca cgtgatccct
     2881 ccagacaaca tcctgcccct tgtcatctgc agcgggatct ccaaccagcc gctggaattc
     2941 acgttccaga agagagagct gcctcagatg atggacgagg tgggtcgcat tctctgtaac
     3001 ctgtgtggtg tggttcctgg aggggtggtc tgtttcttcc cctcctacga gtacctgcgc
     3061 caggtccatg cccactggga gaagggtggc ctgctgggcc gtctggctgc caggaagaag
     3121 atattccagg aacctaagag cgcacaccag gtggagcagg tgctgctggc atattccagg
     3181 tgcatccagg cctgtggcca ggagagaggc caggtgacag gggccctgct cctctctgtg
     3241 gttggaggaa agatgagtga agggatcaac ttctctgaca acctaggccg gtgtgtggtg
     3301 atggtgggca tgcccttccc caacatcagg tctgcagagc tgcaggagaa gatggcctac
     3361 ttggatcaaa ccctccccag agcccccggc caggcacccc cagggaaggc tctggtggag
     3421 aacctgtgca tgaaggccgt caaccagtcc ataggcaggg ccatcaggca ccagaaggat
     3481 tttgccagcg tagtgctcct ggaccagcga tatgcccggc cccctgtcct ggccaagctg
     3541 ccggcctgga tccgagcccg tgtggaggtc aaagctacct ttggccccgc cattgctgct
     3601 gtgcagaagt ttcaccggga gaagtcggcc tcttccgact acaaagacca tgacggtgat
     3661 tataaagatc atgacatcga ttacaaggat gacgatgaca agtgactcga gcatgcatct
     3721 agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt
     3781 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3841 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3901 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3961 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     4021 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     4081 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4141 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4201 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4261 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4321 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4381 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     4441 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     4501 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4561 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4621 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4681 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4741 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4801 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4861 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4921 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4981 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     5041 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     5101 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     5161 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     5221 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5281 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5341 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5401 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5461 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5521 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5581 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5641 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     5701 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5761 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5821 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5881 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5941 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     6001 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     6061 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     6121 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     6181 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6241 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6301 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6361 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6421 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6481 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6541 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6601 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6661 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6721 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6781 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6841 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6901 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6961 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     7021 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     7081 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     7141 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7201 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7261 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7321 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7381 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7441 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7501 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7561 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7621 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7681 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7741 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7801 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7861 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7921 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7981 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     8041 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     8101 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     8161 gaaaagtgcc acctgacg
//
