LOCUS       sequence-217739-        6501 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses a PAR-binding (PBZ) domain fused to mRuby2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6501)
  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 6501)
  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: pcDNA3-PBZ-mRuby2
FEATURES             Location/Qualifiers
     source          1..6501
                     /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     249..268
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        249..267
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             294..317
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             603..632
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="ASMTGGQQMG"
     CDS             join(636..644,645..659)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             666..1376
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     promoter        complement(1440..1458)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1441..1458)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1478..1495)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1484..1708
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1754..2182
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1841..1860)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2051..2072
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2191..2211)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2196..2525
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2376..2511
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2438..2457
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2592..3386
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2646..2665)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3256..3275
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3560..3681
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3597..3616)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3651..3670
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3730..3746)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3730..3746)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3743..3765)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3754..3770
                     /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(3778..3784,3785..3802,3803..3808))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3823..3844
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3961..3978)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4132..4720)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4212..4231)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4891..5682,5683..5751))
                     /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     5514..5533
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5752..5856)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5931..5950)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6122..6501
                     /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 agctctctgg ctaactagag aacccactgc ttactggctt
      241 atcgaaatta atacgactca ctatagggag acccaagctt accggtcacc atggactaca
      301 aagacgatga cgacaagctt gcggccgcga attcaggaaa caaggtcaag aggacatcct
      361 gcatgtatgg ggcaaactgc tataggaaga atcctgttca ttttcaacat tttagccatc
      421 ctggtgatag tgattatgga ggtgttcaaa tcgtgggcca agatgagact gatgaccggc
      481 ctgaatgtcc ctatggacca tcctgttata ggaagaatcc ccagcacaag atagaatata
      541 gacataatac gcttccagtg agaaatgttt tagatgaatc ggtaccagtc gactctagaa
      601 gagctagcat gactggtgga cagcaaatgg gtcgggatct gtacgacgat gacgataagg
      661 atccgatggt gtctaagggc gaagagctga tcaaggaaaa tatgcgtatg aaggtggtca
      721 tggaaggttc ggtcaacggc caccaattca aatgcacagg tgaaggagaa ggcaatccgt
      781 acatgggaac tcaaaccatg aggatcaaag tcatcgaggg aggacccctg ccatttgcct
      841 ttgacattct tgccacgtcg ttcatgtatg gcagccgtac ttttatcaag tacccgaaag
      901 gcattcctga tttctttaaa cagtcctttc ctgagggttt tacttgggaa agagttacga
      961 gatacgaaga tggtggagtc gtcaccgtca tgcaggacac cagccttgag gatggctgtc
     1021 tcgtttacca cgtccaagtc agaggggtaa actttccctc caatggtccc gtgatgcaga
     1081 agaagaccaa gggttgggag cctaatacag agatgatgta tccagcagat ggtggtctga
     1141 ggggatacac tcatatggca ctgaaagttg atggtggtgg ccatctgtct tgctctttcg
     1201 taacaactta caggtcaaaa aagaccgtcg ggaacatcaa gatgcccggt atccatgccg
     1261 ttgatcaccg cctggaaagg ttagaggaaa gtgacaatga aatgttcgta gtacaacgcg
     1321 aacacgcagt tgccaagttc gccgggcttg gtggtgggat ggacgagctg tacaagtaag
     1381 aattctgcag atatccatca cactggcggc cgctcgagca tgcatctaga gggccctatt
     1441 ctatagtgtc acctaaatgc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc
     1501 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     1561 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     1621 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     1681 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     1741 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     1801 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     1861 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     1921 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     1981 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     2041 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     2101 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     2161 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     2221 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     2281 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2341 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     2401 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     2461 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     2521 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     2581 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     2641 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     2701 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     2761 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     2821 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     2881 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     2941 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     3001 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     3061 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     3121 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     3181 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     3241 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     3301 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     3361 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     3421 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     3481 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     3541 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     3601 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     3661 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     3721 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3781 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     3841 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     3901 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3961 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4021 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4081 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4141 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4201 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4261 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4321 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4381 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4441 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4501 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4561 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4621 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4681 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4741 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4801 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4861 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4921 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4981 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5041 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5101 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5161 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5221 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5281 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5341 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5401 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5461 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5521 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5581 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5641 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5701 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5761 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5821 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5881 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
     5941 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     6001 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     6061 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
     6121 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
     6181 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     6241 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     6301 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     6361 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     6421 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     6481 ttagtcatcg ctattaccat g
//
