LOCUS       sequence-440389-        8135 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector of flag-tagged RECQL5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8135)
  AUTHORS   Zhao Y, Tabet D, Rubio Contreras D, Lao L, Kousholt AN, Weile J, 
            Melo H, Hoeg L, Feng S, Cote AG, Lin ZY, Setiaputra D, Jonkers J, 
            Gingras AC, Gomez Herreros F, Roth FP, Durocher D
  TITLE     Genome-scale mapping of DNA damage suppressors through phenotypic 
            CRISPR-Cas9 screens.
  JOURNAL   Mol Cell. 2023 Aug 3;83(15):2792-2809.e9. doi: 
            10.1016/j.molcel.2023.06.025. Epub 2023 Jul 20.
  PUBMED    37478847
REFERENCE   2  (bases 1 to 8135)
  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"; doi: 
            "10.1016/j.molcel.2023.06.025"; journalName: "Mol Cell"; date: 
            "2023-08-3- 3"; volume: "83"; issue: "15"; pages: "2792-2809.e9"
COMMENT     Sequence Label: pcDNA5-FRT/T0-Flag-RECQL5
FEATURES             Location/Qualifiers
     source          1..8135
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        158..537
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        538..741
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     692..712
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    743..761
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    764..782
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     792..816
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             916..939
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4010..4027)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4016..4240
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4373..4392)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4524..4571
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             4579..5601
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    5731..5852
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5768..5787)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5822..5841
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5901..5917)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5901..5917)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5914..5936)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5925..5941
                     /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(5949..5955,5956..5973,5974..5979))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5994..6015
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6132..6149)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6303..6891)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6383..6402)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7062..7853,7854..7922))
                     /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     7685..7704
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7923..8027)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8102..8121)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
       61 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt
      121 gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat
      181 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
      241 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      301 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      361 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      421 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      481 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      541 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      601 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      661 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      721 gaggtctata taagcagagc tctccctatc agtgatagag atctccctat cagtgataga
      781 gatcgtcgac gagctcgttt agtgaaccgt cagatcgcct ggagacgcca tccacgctgt
      841 tttgacctcc atagaagaca ccgggaccga tccagcctcc ggactctagc gtttaaactt
      901 aagcttggta ccatggacta caaagacgat gacgataaag ccgccagttt aaacggcgcg
      961 ccgatgagca gccaccatac cacctttcct tttgaccctg agcggcgagt ccggagtacg
     1021 ctgaagaagg tctttgggtt tgactctttt aagacgcctt tacaggagag tgcgaccatg
     1081 gctgtagtaa aaggtaacaa ggacgtcttt gtgtgcatgc ccacaggggc aggaaaatcc
     1141 ctatgctatc agctccctgc tctgttggcc aaaggcatca ccattgtagt ctctcctctc
     1201 attgctttga ttcaggacca agtggaccac ttgctaaccc taaaggtacg agtaagttcc
     1261 ctgaactcga agctctctgc acaggaaagg aaggagctgc ttgctgacct ggagcgagaa
     1321 aagccccaga ccaagattct gtacatcacc ccagagatgg cagcttcatc ctccttccag
     1381 cccaccctga actccctggt gtcccgccac ctgctgtctt acttggtggt ggatgaagct
     1441 cattgtgttt cccaatgggg gcatgacttt cgtcctgact acttgcgtct gggtgccctg
     1501 cgctcccgcc tgggacatgc cccttgtgtg gctctgaccg ccacagccac cccacaggtc
     1561 caagaggacg tgtttgctgc cctgcacctg aagaaaccag ttgccatctt caagactccc
     1621 tgcttccggg ccaacctctt ctatgatgtg caattcaagg aactgatttc tgatccctat
     1681 gggaacctga aggacttctg ccttaaggct cttggacagg aggctgataa agggttatct
     1741 ggctgcggca ttgtgtactg caggactaga gaggcttgtg aacagctggc catagagctc
     1801 agctgcaggg gtgtgaacgc caaggcttac catgcagggc tgaaggcctc tgaaagaacg
     1861 ctggtgcaga acgactggat ggaggagaag gtccctgtaa ttgttgcaac cattagtttt
     1921 gggatgggag tggataaagc caatgtcagg tttgtcgccc attggaatat tgccaagtct
     1981 atggctgggt actaccagga gtctggccgg gctggcaggg atgggaagcc ttcctggtgc
     2041 cgtctctatt actccaggaa tgaccgggac caagtcagct tcctgatcag gaaggaagta
     2101 gcaaaactcc aggaaaagag aggaaacaaa gcatctgata aagccactat catggccttt
     2161 gatgccctgg tgaccttctg tgaagaactg gggtgccgcc atgccgccat tgccaagtac
     2221 ttcggggatg cgctgcctgc ctgcgccaaa ggctgcgacc actgccagaa ccccacggcc
     2281 gtgcggaggc ggctggaggc cttggagcgc agcagcagct ggagcaagac ctgcatcggg
     2341 ccctcccagg ggaacggctt tgaccccgag ctgtacgagg gaggccgcaa gggctacggg
     2401 gacttcagca ggtatgacga aggttctgga ggcagcgggg atgaaggcag agatgaggcc
     2461 cacaagcggg agtggaacct cttctatcag aagcagatgc agctgcgcaa gggcaaagac
     2521 cccaagatag aagaatttgt acccccagat gagaactgtc ccctgaaaga ggcttctagc
     2581 aggaggatcc ccaggctgac tgtgaaggca cgggagcact gcctgcggct tctggaggag
     2641 gcgctgagca gcaaccgcca gtcaacacgt accgctgatg aagctgacct ccgggccaag
     2701 gccgtggagc tggaacatga gacattccgg aacgccaagg tggccaacct ctacaaggcc
     2761 agcgtgctga agaaggtggc cgatatccac agagcctcca aggatgggca gccctatgac
     2821 atgggaggca gtgccaagag ctgcagtgcc caagctgagc ccccggagcc caatgagtat
     2881 gacattccac cagcctccca tgtgtactcg ctcaaaccca agcgggtggg agctggtttc
     2941 cccaaaggct cctgcccgtt ccagacggcc acggaactga tggagacgac tcggatcagg
     3001 gagcaagccc cccagcccga gcggggaggc gagcatgagc ccccgagccg gccctgtggc
     3061 ctcctggatg aggatgggag tgagcccctc cctgggccca gaggggaggt ccctggaggc
     3121 agcgctcact atggggggcc ctcccctgag aagaaggcaa aaagttcctc tgggggcagc
     3181 tcccttgcca agggccgggc tagcaagaaa cagcagctcc tagccacagc ggcccacaag
     3241 gattctcaga gcatcgcccg cttcttctgc cgaagggtgg aaagcccagc tctgctggca
     3301 tcagccccag aggcagaagg tgcctgcccc tcctgtgagg gggttcaggg acccccgatg
     3361 gccccagaga agtacacagg ggaggaagat ggagccgggg gacattcgcc tgcccctccc
     3421 cagactgagg agtgcctcag ggagaggcca agcacctgcc cgcccagaga ccagggcacc
     3481 cctgaagtcc agcccacccc tgcaaaggac acatggaagg gcaagcggcc tcgatcccag
     3541 caggagaacc cagagagcca gcctcagaag aggccacgcc cctcagccaa gccctccgtc
     3601 gtagctgagg tcaagggcag cgtctcggcc agcgaacagg gcaccttgaa tcccacggct
     3661 caagacccct tccagctctc cgctcctggc gtctccttga aggaggctgc aaatgttgtg
     3721 gtcaagtgcc tcaccccttt ctacaaggag ggcaagtttg cttccaagga gttgtttaaa
     3781 ggctttgccc gccacctctc acacttgctg actcagaaga cctctcctgg aaggagcgtg
     3841 aaagaagagg cccagaacct catcaggcac ttcttccatg gccgggcccg gtgcgagagc
     3901 gaagctgact ggcatggcct gtgtggcccc cagagatgag aattctgcag atatccatca
     3961 cactggcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg
     4021 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     4081 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     4141 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     4201 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     4261 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4321 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4381 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4441 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4501 tagggtgatg gttcacgtac ctagaagttc ctattccgaa gttcctattc tctagaaagt
     4561 ataggaactt ccttggccaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg
     4621 atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt
     4681 gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat
     4741 ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg
     4801 gaagtgcttg acattgggga gttcagcgag agcctgacct attgcatctc ccgccgtgca
     4861 cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc
     4921 gcggaggcca tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca
     4981 ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct
     5041 gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg
     5101 caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg
     5161 cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt
     5221 gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg
     5281 aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag
     5341 cttgcaggat cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat
     5401 cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca
     5461 atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc
     5521 gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact
     5581 cgtccgaggg caaaggaata gcacgtacta cgagatttcg attccaccgc cgccttctat
     5641 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5701 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     5761 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5821 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     5881 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     5941 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     6001 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     6061 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     6121 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     6181 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     6241 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     6301 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6361 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6421 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6481 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6541 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6601 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6661 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6721 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     6781 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6841 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6901 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6961 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     7021 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     7081 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     7141 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     7201 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     7261 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     7321 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     7381 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     7441 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     7501 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7561 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7621 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7681 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7741 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7801 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7861 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7921 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7981 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     8041 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca
     8101 gtacaatctg ctctgatgcc gcatagttaa gccag
//
