LOCUS       sequence-440388-        8896 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector of flag-tagged BRIP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8896)
  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 8896)
  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-BRIP1
FEATURES             Location/Qualifiers
     source          1..8896
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        168..547
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        548..751
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     702..722
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    753..771
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    774..792
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     802..826
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             926..949
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4781..4798)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4787..5011
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5144..5163)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    5295..5342
                     /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             5350..6372
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    6502..6623
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6539..6558)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6593..6612
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6672..6688)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6672..6688)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6685..6707)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6696..6712
                     /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(6720..6726,6727..6744,6745..6750))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6765..6786
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6903..6920)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7074..7662)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7154..7173)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7833..8624,8625..8693))
                     /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     8456..8475
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8694..8798)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8873..8892)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
       61 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
      121 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt
      181 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      241 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      301 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      361 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      421 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      481 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      541 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      601 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      661 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      721 tgtacggtgg gaggtctata taagcagagc tctccctatc agtgatagag atctccctat
      781 cagtgataga gatcgtcgac gagctcgttt agtgaaccgt cagatcgcct ggagacgcca
      841 tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc ggactctagc
      901 gtttaaactt aagcttggta ccatggacta caaagacgat gacgataaag ccgccagttt
      961 aaacggcgcg ccgtcttcaa tgtggtctga atatacaatt ggtggggtga agatttactt
     1021 tccttataaa gcttacccgt cacagcttgc tatgatgaat tctattctca gaggattaaa
     1081 cagcaagcaa cattgtttgt tggagagtcc cacaggaagt ggaaaaagct tagccttact
     1141 ttgttctgct ttagcatggc aacaatctct tagtgggaaa ccagcagatg agggcgtaag
     1201 tgaaaaagct gaagtacaat tgtcatgttg ttgtgcatgc cattcaaagg attttacaaa
     1261 caatgacatg aaccaaggaa cttcacgtca tttcaactat ccaagcacac caccttctga
     1321 aagaaatggc acttcatcaa cttgtcaaga ctcccctgaa aaaaccactc tggctgcaaa
     1381 gttatctgct aagaaacagg catccatata cagagatgaa aatgatgatt ttcaagtaga
     1441 gaagaaaaga attcgaccct tagaaactac acagcagatt agaaaacgtc attgctttgg
     1501 aacagaagta cacaatttgg atgcaaaagt tgattcagga aagactgtaa aactcaactc
     1561 tccactggaa aagataaact ccttttcgcc acagaaaccc cctggccact gttctaggtg
     1621 ctgttgttct actaaacaag gaaacagtca agagtcatcg aataccatta agaaggatca
     1681 tacagggaaa tccaagatac ccaaaatata ttttgggaca cgcacacaca agcagattgc
     1741 tcagattact agagagctcc ggaggacggc atattcaggg gttccaatga ctattctttc
     1801 cagcagggat catacttgtg tccatcctga ggtagtcggt aacttcaaca gaaatgagaa
     1861 gtgcatggaa ttgctagatg ggaaaaacgg aaaatcctgc tatttttatc atggagttca
     1921 taaaattagt gatcagcaca cattacagac tttccaaggg atgtgcaaag cctgggatat
     1981 agaagaactt gtcagcctgg ggaagaaact aaaggcctgt ccatattaca cagcccgaga
     2041 actaatacaa gatgctgaca tcatattttg tccctacaac tatcttctag atgcacaaat
     2101 aagggaaagt atggatttaa atctgaaaga acaggttgtc attttagatg aagctcataa
     2161 catcgaggac tgtgctcggg aatcagcaag ttacagtgta acagaagttc agcttcggtt
     2221 tgctcgggat gaactagata gtatggtcaa caataatata aggaagaaag atcatgaacc
     2281 cctacgagct gtgtgctgta gcctcattaa ttggttagaa gcaaacgctg aatatcttgt
     2341 agaaagagat tatgaatcag cttgtaaaat atggagtgga aatgaaatgc tcttaacttt
     2401 acacaaaatg ggtatcacca ctgctacttt tcccattttg cagggacatt tttctgctgt
     2461 tcttcaaaaa gaggaaaaaa tctcaccaat ttatggtaaa gaggaggcaa gagaagtacc
     2521 tgttattagt gcatcaactc aaataatgct taaaggactt tttatggtac ttgactatct
     2581 ttttaggcaa aatagcagat ttgcagatga ttataaaatt gcgattcaac agacttactc
     2641 ctggacaaat cagattgata tttcagacaa aaatgggttg ttggttctac caaaaaataa
     2701 gaaacgttca cgacagaaaa ctgcagttca tgtgctaaac ttttggtgct taaatccagc
     2761 tgtggccttt tcagatatta atggcaaagt tcagaccatt gttttgacat ctggtacatt
     2821 atcaccaatg aaatcctttt cgtcagaact tggtgttaca tttactatcc agctggaggc
     2881 taatcatatc attaaaaatt cacaggtttg ggttggtacc attgggtcag gccccaaggg
     2941 tcggaatctc tgtgctacct tccagaatac tgaaacattt gagttccaag atgaagtggg
     3001 agcacttttg ttatctgtgt gccagactgt gagccaagga attttgtgtt tcttgccatc
     3061 ttacaagtta ttagaaaaat taaaagaacg ttggctctct actggtttat ggcataatct
     3121 ggagttggtg aagacagtca ttgtagaacc acagggagga gaaaaaacaa attttgatga
     3181 attactgcag gtgtactatg acgcaatcaa atacaaagga gagaaagatg gagctctcct
     3241 ggtagcagtt tgtcgtggta aagtgagtga gggtctggat ttctcagatg acaatgcccg
     3301 tgctgtcata acaataggaa ttccttttcc aaatgtgaaa gatctacagg ttgaactaaa
     3361 acgacaatac aatgaccacc attcaaaatt gagaggtctt ctacctggcc gtcagtggta
     3421 tgaaattcaa gcatacaggg ccttaaacca ggcccttggt agatgtatta gacacagaaa
     3481 tgattgggga gctcttattc tagtggatga tcgctttagg aataacccaa gtcgctatat
     3541 atctggactt tctaaatggg tacggcagca gattcagcac cattcaacct ttgaaagtgc
     3601 actggagtcc ttggctgaat tttccaaaaa gcatcaaaaa gttcttaatg tatccataaa
     3661 ggacagaacc aatatacagg acaatgagtc tacacttgaa gtgacctctt taaagtacag
     3721 taccccacct tatttactgg aagcagcaag tcatctatca ccagaaaatt ttgtggaaga
     3781 tgaagcaaag atatgtgtcc aggaactaca gtgtcctaaa attattacca aaaattcacc
     3841 tctaccaagt agcattatct ccagaaagga gaaaaatgat ccagtattcc tggaagaagc
     3901 agggaaagca gaaaaaattg tgatttccag atccacaagc ccaactttca acaaacaaac
     3961 aaagagagtt agctggtcaa gctttaattc tttgggacag tattttactg gtaaaatacc
     4021 gaaggcaaca cctgagctcg ggtcatcaga gaatagtgcc tctagtcctc cccgtttcaa
     4081 aacagagaag atggaaagta aaactgtttt gcccttcact gataaatgtg aatcctcaaa
     4141 tctgacagta aacacatcgt ttggatcatg ccctcaatca gaaaccatta tttcatcatt
     4201 aaagattgat gccaccctta ctagaaaaaa tcattctgaa catccgctct gttctgaaga
     4261 agccctggat ccagacattg aattgtctct agtaagtgaa gaagataaac agtccacttc
     4321 aaatagagat tttgaaacag aagcagaaga tgaatctatc tattttacac ctgaacttta
     4381 cgatcctgaa gatacagatg aagaaaaaaa tgacctagct gaaactgata gaggaaatag
     4441 attggctaac aattcagatt gcattttagc taaagacctt tttgaaatta gaactataaa
     4501 agaagtagat tcagccagag aagtgaaagc tgaggattgc atagatacaa agttgaatgg
     4561 aattctgcat attgaagaaa gtaaaattga tgacattgat ggtaatgtaa aaacaacttg
     4621 gataaatgaa ctggaactgg gaaaaactca tgaaatagaa ataaagaact ttaaaccatc
     4681 tccttccaaa aataaaggca tgtttcctgg ttttaagtga gatatccatc acactggcgg
     4741 ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     4801 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     4861 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     4921 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     4981 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     5041 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     5101 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     5161 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     5221 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     5281 ggttcacgta cctagaagtt cctattccga agttcctatt ctctagaaag tataggaact
     5341 tccttggcca aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag
     5401 ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc
     5461 ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac
     5521 aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt
     5581 gacattgggg agttcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc
     5641 acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc
     5701 atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg
     5761 caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat
     5821 gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc
     5881 gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat
     5941 ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc
     6001 gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg
     6061 ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga
     6121 tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg
     6181 gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga
     6241 tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc
     6301 gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg
     6361 gcaaaggaat agcacgtact acgagatttc gattccaccg ccgccttcta tgaaaggttg
     6421 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     6481 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     6541 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     6601 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     6661 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     6721 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     6781 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     6841 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     6901 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     6961 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     7021 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     7081 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     7141 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     7201 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     7261 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     7321 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     7381 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     7441 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     7501 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     7561 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     7621 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     7681 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     7741 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     7801 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     7861 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     7921 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     7981 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     8041 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     8101 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     8161 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     8221 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     8281 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     8341 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     8401 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     8461 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     8521 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     8581 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     8641 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     8701 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     8761 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     8821 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     8881 gctctgatgc cgcata
//
