LOCUS       sequence-413521-        7601 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for generating Flp-In cell lines allowing dox-inducible 
            mammalian expression of 3xFlag-tagged full length TAF5 isoform.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7601)
  AUTHORS   Xiao MS, Damodaran AP, Kumari B, Dickson E, Xing K, On TA, Parab N, 
            King HE, Perez AR, Guiblet WM, Duncan G, Che A, Chari R, Andresson 
            T, Vidigal JA, Weatheritt RJ, Aregger M, Gonatopoulos-Pournatzis T
  TITLE     Genome-scale exon perturbation screens uncover exons critical for 
            cell fitness.
  JOURNAL   Mol Cell. 2024 Jul 11;84(13):2553-2572.e19. doi: 
            10.1016/j.molcel.2024.05.024. Epub 2024 Jun 24.
  PUBMED    38917794
REFERENCE   2  (bases 1 to 7601)
  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.2024.05.024"; journalName: "Mol Cell"; date: 
            "2024-07-11- 11"; volume: "84"; issue: "13"; pages: "2553-2572.e19"
COMMENT     Sequence Label: pcDNA5-3xFLAG-TAF5-FL
FEATURES             Location/Qualifiers
     source          1..7601
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1034..1057
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    1058..1082
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3487..3511)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3553..3570)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3559..3783
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3916..3935)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4067..4114
                     /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             4122..5144
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    5274..5395
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5311..5330)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5365..5384
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5444..5460)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5444..5460)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5457..5479)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5468..5484
                     /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(5492..5498,5499..5516,5517..5522))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5537..5558
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5675..5692)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5846..6434)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5926..5945)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6605..7396,7397..7465))
                     /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     7228..7247
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7466..7570)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttccaccat ggactacaaa gaccatgacg gtgatacata
     1021 tgatcatgac atcgattaca aggatgacga tgacaagaca agtttgtaca aaaaagcagg
     1081 cttcgcggcg ctggcggagg agcagacgga ggtggcggtc aagctagagc ctgagggacc
     1141 gccaacgctg ctacctccgc aggcggggga cggcgcaggc gagggtagcg gcggcactac
     1201 caacaacggc cccaacggcg gcggcgggaa cgttgcggcg tcgtcgtcca ctggcgggga
     1261 tggcgggacc cccaagccca cggtggctgt ctccgccgct gccccggcgg gggcggcccc
     1321 ggtgcccgcc gctgctccgg acgccggcgc tccgcatgac cgacagactc tactggccgt
     1381 gctgcagttc ctacggcaga gcaaactccg cgaggccgaa gaggcgctgc gccgtgaggc
     1441 cgggctgctg gaggaggcag tggcgggctc cggagccccg ggagaggtgg acagcgccgg
     1501 cgctgaggtg accagcgcgc ttctcagccg ggtgaccgcc tcggcccctg gccctgcggc
     1561 ccccgaccct ccgggcactg gcgcttcggg ggccacggtc gtctcaggtt cagcctcagg
     1621 tcctgcggct ccgggtaaag ttggaagtgt tgctgtggaa gaccagccag atgtcagtgc
     1681 cgtgttgtca gcctacaacc aacaaggaga tcccacaatg tatgaagaat actatagtgg
     1741 actgaaacac ttcattgaat gttccctgga ctgccatcgg gcagagttgt cccaactttt
     1801 ttatcctctg tttgtgcaca tgtacttgga gctagtctac aatcaacatg agaatgaagc
     1861 aaagtcattc tttgagaagt tccatggaga tcaggaatgt tattaccagg atgacctacg
     1921 agtattatct agtcttacca aaaaggaaca catgaaaggg aatgagacca tgttggattt
     1981 tcgaacaagt aaatttgttc tgcgtatttc ccgtgactcg taccaactct tgaagaggca
     2041 tcttcaggag aaacagaaca atcagatatg gaacatagtt caggagcacc tctacattga
     2101 catctttgat gggatgccgc gtagtaagca acagatagat gcgatggtgg gaagtttggc
     2161 aggagaggct aaacgagagg caaacaaatc aaaggtattt tttggtttat taaaagaacc
     2221 agaaattgag gtacctttgg atgacgagga tgaagaggga gaaaatgaag aaggaaaacc
     2281 taaaaagaag aagcctaaaa aagatagtat tggatccaaa agcaaaaaac aagatcccaa
     2341 tgctccacct cagaacagaa tccctcttcc tgagttgaaa gattcagata agttggataa
     2401 gataatgaat atgaaagaaa ccaccaaacg agtgcgcctt gggccggact gcttaccctc
     2461 catttgtttc tatacatttc tcaatgctta ccagggtctc actgcagtgg atgtcactga
     2521 tgattctagt ctgattgctg gaggttttgc agattcaact gtcagagtgt ggtcggtaac
     2581 acccaaaaag cttcgtagtg tcaaacaagc atcagatctt agtcttatag acaaagaatc
     2641 agatgatgtc ttagaaagaa tcatggatga gaaaacagca agtgagttga agattttgta
     2701 tggtcacagt gggcctgtct acggagccag cttcagtccg gataggaact atctgctttc
     2761 ctcttcagag gacggaactg ttagattgtg gagccttcaa acatttactt gtttggtggg
     2821 atataaagga cacaactatc cagtatggga cacacaattt tctccatatg gatattattt
     2881 tgtgtcaggg ggccatgacc gagtagctcg gctctgggct acagaccact atcagccttt
     2941 aagaatattt gccggccatc ttgctgatgt gaattgtacc agattccatc caaattctaa
     3001 ttatgttgct acgggctctg cagacagaac tgtgcggctc tgggacgtcc tgaatggtaa
     3061 ctgtgtaagg atcttcactg gacacaaggg accaattcat tccttgacat tttctcccaa
     3121 tgggagattc ctggctacag gagcaacaga tggcagagtg cttctttggg atattggaca
     3181 tggtttgatg gttggagaat taaaaggcca cactgataca gtctgttcac ttaggtttag
     3241 tagagatggt gaaattttgg catcaggttc aatggataat acagttcgat tatgggatgc
     3301 tatcaaagcc tttgaagatt tagagaccga tgactttact acagccactg ggcatataaa
     3361 tttacctgag aattcacagg agttattgtt gggaacatat atgaccaaat caacaccagt
     3421 tgtacacctt cattttactc gaagaaacct ggttctagct gcaggagctt atagtccaca
     3481 atgataaccc agctttcttg tacaaagtgg tctgactcga gtctagaggg cccgtttaaa
     3541 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     3601 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     3661 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     3721 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     3781 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     3841 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3901 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3961 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4021 acctcgaccc caaaaaactt gattagggtg atggttcacg tacctagaag ttcctattcc
     4081 gaagttccta ttctctagaa agtataggaa cttccttggc caaaaagcct gaactcaccg
     4141 cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc
     4201 tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc
     4261 tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg
     4321 catcggccgc gctcccgatt ccggaagtgc ttgacattgg ggagttcagc gagagcctga
     4381 cctattgcat ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac
     4441 tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg gccgatctta
     4501 gccagacgag cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc
     4561 gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg
     4621 acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact
     4681 gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca
     4741 atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg
     4801 aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct
     4861 acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg tatatgctcc
     4921 gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt
     4981 gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac
     5041 aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta ctcgccgata
     5101 gtggaaaccg acgccccagc actcgtccga gggcaaagga atagcacgta ctacgagatt
     5161 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5221 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     5281 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5341 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5401 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     5461 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5521 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5581 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5641 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5701 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5761 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5821 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5881 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5941 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6001 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6061 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6121 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6181 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6241 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6301 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6361 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6421 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6481 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6541 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6601 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6661 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6721 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6781 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6841 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6901 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6961 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7021 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7081 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7141 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7201 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7261 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7321 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7381 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7441 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7501 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7561 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
