LOCUS       sequence-489574-        5649 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for generating Flp-In cell lines allowing dox-inducible 
            mammalian expresion of ZNF207-ZnF+Hel_K42M_3xFLAG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5649)
  AUTHORS   Behera AK, Kim JJ, Kordale S, Damodaran AP, Kumari B, Vidak S, 
            Dickson E, Xiao MS, Duncan G, Andresson T, Misteli T, 
            Gonatopoulos-Pournatzis T
  TITLE     RNA-coupled CRISPR Screens Reveal ZNF207 as a Regulator of LMNA 
            Aberrant Splicing in Progeria.
  JOURNAL   bioRxiv [Preprint]. 2025 Apr 26:2025.04.25.648738. doi: 
            10.1101/2025.04.25.648738.
  PUBMED    40568141
REFERENCE   2  (bases 1 to 5649)
  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: "bioRxiv 
            [Preprint]. 2025 Apr 26:2025.04.25.648738. doi: 
            10.1101/2025.04.25.648738."
COMMENT     Sequence Label: pcDNA5-ZNF207-ZnF+Hel_K42M_3xFLAG
FEATURES             Location/Qualifiers
     source          1..5649
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    390..511
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(427..446)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     481..500
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(560..576)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(560..576)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(573..595)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    584..600
                     /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(608..614,615..632,633..638))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    653..674
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(791..808)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(962..1550)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1042..1061)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1721..2512,2513..2581))
                     /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     2344..2363
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2582..2686)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2761..2780)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2952..3331
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3332..3535
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3486..3506
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    3537..3555
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3558..3576
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3586..3610
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    3753..3777
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4147..4171)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4212..4277
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(4318..4335)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4324..4548
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4681..4700)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4832..4879
                     /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             join(4887..5649,1..260)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             join(4887..5649,1..260)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 ttgcaggatc gccgcggctc cgggcgtata tgctccgcat tggtcttgac caactctatc
       61 agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa
      121 tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg
      181 tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc
      241 gtccgagggc aaaggaatag cacgtactac gagatttcga ttccaccgcc gccttctatg
      301 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
      361 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
      421 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
      481 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
      541 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
      601 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
      661 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
      721 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
      781 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
      841 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      901 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      961 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     1021 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1081 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1141 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     1201 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     1261 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     1321 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     1381 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     1441 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     1501 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     1561 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     1621 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     1681 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     1741 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1801 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1861 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1921 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1981 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     2041 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     2101 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     2161 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     2221 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     2281 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     2341 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     2401 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     2461 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     2521 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     2581 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     2641 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     2701 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     2761 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     2821 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     2881 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
     2941 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     3001 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     3061 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     3121 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     3181 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     3241 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     3301 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     3361 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     3421 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     3481 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctccc
     3541 tatcagtgat agagatctcc ctatcagtga tagagatcgt cgacgagctc gtttagtgaa
     3601 ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga
     3661 ccgatccagc ctccggactc tagcgtttaa acttaagctt ggtaccgagc tcggatccac
     3721 tagtccagtg tggtggaatt ctgcagatat caacaagttt gtacaaaaaa gcaggcttca
     3781 tgggtcgcaa gaagaagaag cagctgaagc cgtggtgctg gtattgtaat agagattttg
     3841 atgatgagaa gatccttatt cagcaccaaa aagcaaagca ttttaaatgc catatatgtc
     3901 acatgaaatt gtatacagga cctggcttag ctattcattg catgcaggta cataaagaaa
     3961 caatagatgc cgtaccaaat gcaatacctg gaagaacaga catagagttg gaaatatatg
     4021 gtatggaagg tattccagaa aaagacatgg atgaaagacg acgacttctt gaacagaaaa
     4081 cacaagaaag tcaaaaaaag aagcaacaag atgattctga tgaatatgat gatgacgact
     4141 ctgcaaaccc agctttcttg tacaaagtgg tgatgggctg caggaattcg atatcaagct
     4201 taggtggaat ggactacaag gaccacgacg gtgactacaa ggaccacgac atcgactaca
     4261 aggacgacga cgacaagtag ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct
     4321 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     4381 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     4441 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     4501 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     4561 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     4621 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     4681 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4741 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4801 aacttgatta gggtgatggt tcacgtacct agaagttcct attccgaagt tcctattctc
     4861 tagaaagtat aggaacttcc ttggccaaaa agcctgaact caccgcgacg tctgtcgaga
     4921 agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag
     4981 aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct
     5041 gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc
     5101 cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc
     5161 gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc
     5221 agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt
     5281 tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg
     5341 cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt
     5401 ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc
     5461 acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag
     5521 cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct
     5581 tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc
     5641 atccggagc
//
