LOCUS       sequence-208101-        4313 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses a PAR-binding (PBZ) domain fused to venus C-terminus, 
            contains point mutations that abolish PAR binding.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4313)
  AUTHORS   Krastev DB, Pettitt SJ, Campbell J, Song F, Tanos BE, Stoynov SS, 
            Ashworth A, Lord CJ
  TITLE     Coupling bimolecular PARylation biosensors with genetic screens to 
            identify PARylation targets.
  JOURNAL   Nat Commun. 2018 May 22;9(1):2016. doi: 10.1038/s41467-018-04466-4.
  PUBMED    29789535
REFERENCE   2  (bases 1 to 4313)
  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: "Nat 
            Commun."; date: "2018-05-22"; volume: "9(1):2016. doi"; pages: " 
            10.1038/s41467-018-04466-4"
COMMENT     Sequence Label: pBiFC-PBZ-4A-VC
FEATURES             Location/Qualifiers
     source          1..4313
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4..30
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             358..609
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
                     YLSYQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     546..567
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    624..758
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(648..667)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     702..721
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(869..885)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(869..885)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(882..904)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    893..909
                     /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(917..923,924..941,942..947))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    962..983
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1100..1117)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1271..1859)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1351..1370)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2030..2821,2822..2890))
                     /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     2653..2672
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2891..2995)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3063..3081
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3119..3141)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3241..3260
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3454..3476
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3468..3485
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3469..3485
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        3512..3815
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3816..4019
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3970..3990
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4016..4040
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          4157..4253
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
ORIGIN
        1 atgtacccat acgatgttcc agattacgct cttatggcca tggaggcccg aattcgggga
       61 aacaaggtca agaggacatc cgccatgtat ggggcaaacg cctataggaa gaatcctgtt
      121 cattttcaac attttagcca tcctggtgat agtgattatg gaggtgttca aatcgtgggc
      181 caagatgaga ctgatgaccg gcctgaagcc ccctatggac catccgccta taggaagaat
      241 ccccagcaca agatagaata tagacataat acgcttccag tgagaaatgt tttagatgaa
      301 ggtacccgtc cggcgtgcaa aatcccgaac gacctgaaac agaaagtcat gaaccacgac
      361 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc
      421 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
      481 cccgacaacc actacctgag ctaccagtcc aaactgagca aagaccccaa cgagaagcgc
      541 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
      601 ctgtacaagt aagcggccgc ggggatccag acatgataag atacattgat gagtttggac
      661 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
      721 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
      781 ttatgtttca ggttcagggg gaggtgtggg aggttttttc ggatcctcta gagtcgatct
      841 gcaggcatgc tagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
      901 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
      961 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     1021 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     1081 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     1141 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     1201 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     1261 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     1321 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1381 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     1441 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     1501 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     1561 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     1621 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     1681 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     1741 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     1801 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     1861 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     1921 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     1981 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     2041 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     2101 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     2161 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     2221 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     2281 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     2341 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     2401 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     2461 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     2521 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     2581 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     2641 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     2701 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     2761 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     2821 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     2881 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     2941 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     3001 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     3061 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     3121 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     3181 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     3241 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
     3301 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg
     3361 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     3421 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     3481 ccagtgagtt cgagcttgca tgcctgcagg tcgttacata acttacggta aatggcccgc
     3541 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     3601 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     3661 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     3721 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     3781 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     3841 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     3901 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     3961 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
     4021 gtttagtgaa ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa
     4081 gacaccggga ccgatccagc ctccggactc tagaggatcc ggtactagag gaactgaaaa
     4141 accagaaagt taactggtaa gtttagtctt tttgtctttt atttcaggtc ccggatccgg
     4201 tggtggtgca aatcaaagaa ctgctcctca gtggatgttg cctttacttc taggcctgta
     4261 cggaagtgtt acttctgctc taaaagctgc ggaattgtac ccgcgggccc acc
//
