LOCUS       sequence-217742-        4313 bp    DNA     circular SYN 14-APR-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 Apr 14, 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"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     201..225
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          342..438
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             502..528
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             856..1107
                     /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     1044..1065
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1122..1256
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1146..1165)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1200..1219
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1367..1383)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1367..1383)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1380..1402)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1391..1407
                     /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(1415..1421,1422..1439,1440..1445))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1460..1481
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1598..1615)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1769..2357)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1849..1868)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2528..3319,3320..3388))
                     /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     3151..3170
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3389..3493)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3561..3579
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3617..3639)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3739..3758
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3952..3974
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3966..3983
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3967..3983
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4010..4313
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
      241 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg gactctagag
      301 gatccggtac tagaggaact gaaaaaccag aaagttaact ggtaagttta gtctttttgt
      361 cttttatttc aggtcccgga tccggtggtg gtgcaaatca aagaactgct cctcagtgga
      421 tgttgccttt acttctaggc ctgtacggaa gtgttacttc tgctctaaaa gctgcggaat
      481 tgtacccgcg ggcccaccat gtacccatac gatgttccag attacgctct tatggccatg
      541 gaggcccgaa ttcggggaaa caaggtcaag aggacatccg ccatgtatgg ggcaaacgcc
      601 tataggaaga atcctgttca ttttcaacat tttagccatc ctggtgatag tgattatgga
      661 ggtgttcaaa tcgtgggcca agatgagact gatgaccggc ctgaagcccc ctatggacca
      721 tccgcctata ggaagaatcc ccagcacaag atagaatata gacataatac gcttccagtg
      781 agaaatgttt tagatgaagg tacccgtccg gcgtgcaaaa tcccgaacga cctgaaacag
      841 aaagtcatga accacgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac
      901 aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
      961 gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa actgagcaaa
     1021 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     1081 actctcggca tggacgagct gtacaagtaa gcggccgcgg ggatccagac atgataagat
     1141 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     1201 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     1261 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttcgg
     1321 atcctctaga gtcgatctgc aggcatgcta gcttggcgta atcatggtca tagctgtttc
     1381 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     1441 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     1501 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     1561 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     1621 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     1681 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     1741 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     1801 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     1861 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     1921 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1981 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2041 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2101 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2161 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2221 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2281 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2341 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2401 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     2461 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     2521 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     2581 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     2641 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     2701 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     2761 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     2821 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     2881 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     2941 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     3001 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     3061 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3121 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     3181 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     3241 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     3301 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     3361 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     3421 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     3481 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     3541 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     3601 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     3661 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     3721 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     3781 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca
     3841 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     3901 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     3961 gacgttgtaa aacgacggcc agtgagttcg agcttgcatg cctgcaggtc gttacataac
     4021 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     4081 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     4141 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     4201 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     4261 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atg
//
