LOCUS       sequence-208095-        5444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses a PAR-binding (PBZ) domain fused to venus N-terminus, 
            contains point mutations that abolish PAR binding.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5444)
  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 5444)
  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-VN
FEATURES             Location/Qualifiers
     source          1..5444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             935..958
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      1250..1259
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1256..1258,1259..1261,1262..1774)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(1301..1322)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1562..1581)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    1785..2407
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1984..2003)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2431..2451)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2436..2765
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2616..2751
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2803..2822)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2804..2822)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2836..2852)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2836..2852)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2849..2871)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2860..2876
                     /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(2884..2890,2891..2908,2909..2914))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2929..2950
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3067..3084)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3238..3826)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3318..3337)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3997..4788,4789..4857))
                     /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     4620..4639
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4858..4962)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4989..5417
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5076..5095)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5286..5307
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attgatctac catggactac aaagacgatg acgacaagct
      961 tgcggccgcg aattcaggaa acaaggtcaa gaggacatcc gccatgtatg gggcaaacgc
     1021 ctataggaag aatcctgttc attttcaaca ttttagccat cctggtgata gtgattatgg
     1081 aggtgttcaa atcgtgggcc aagatgagac tgatgaccgg cctgaagccc cctatggacc
     1141 atccgcctat aggaagaatc cccagcacaa gatagaatat agacataata cgcttccagt
     1201 gagaaatgtt ttagatgaat cggtaccagt cgactctaga agatccatcg ccaccatggt
     1261 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1321 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     1381 gctgaccctg aagctgatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     1441 gaccaccctg ggctacggcc tgcagtgctt cgcccgctac cccgaccaca tgaagcagca
     1501 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1561 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1621 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1681 ggagtacaac tacaacagcc acaacgtcta tatcaccgcc gacaagcaga agaacggcat
     1741 caaggccaac ttcaagatcc gccacaacat cgagtaggga tcccgggtgg catccctgtg
     1801 acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg
     1861 tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg
     1921 ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg
     1981 gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg
     2041 cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat
     2101 gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg
     2161 ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc
     2221 tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgatttta aaataactat
     2281 accagcagga ggacgtccag acacagcata ggctacctgg ccatgcccaa ccggtgggac
     2341 atttgagttg cttgcttggc actgtcctct catgcgttgg gtccactcag tagatgcctg
     2401 ttgaattggg tacgcggcca gcttggctgt ggaatgtgtg tcagttaggg tgtggaaagt
     2461 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     2521 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2581 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     2641 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     2701 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     2761 gcaaaaagct cctcgaggaa ctgaaaaacc agaaagttaa ttccctatag tgagtcgtat
     2821 taaattcgta atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     2881 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     2941 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3001 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3061 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3121 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3181 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3241 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3301 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3361 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3421 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3481 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3541 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3601 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3661 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3721 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3781 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3841 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3901 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3961 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4021 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4081 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4141 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4201 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4261 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4321 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4381 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4441 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4501 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4561 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4621 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4681 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4741 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4801 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4861 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4921 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4981 gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     5041 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     5101 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     5161 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     5221 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     5281 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     5341 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     5401 aaaatttaac gcgaatttta acaaaatatt aacgcttaca attt
//
