LOCUS       sequence-208102-        6502 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 6502)
  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 6502)
  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..6502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             908..931
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1217..1246
                     /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(1250..1258,1259..1273)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1280..1990
                     /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(2054..2072)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2055..2072)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2092..2109)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2098..2322
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2368..2796
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2455..2474)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2665..2686
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2805..2825)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2810..3140
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2991..3126
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3053..3072
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3207..4001
                     /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(3261..3280)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3871..3890
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4175..4296
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4212..4231)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4266..4285
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4345..4361)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4345..4361)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4358..4380)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4369..4385
                     /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(4393..4399,4400..4417,4418..4423))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4438..4459
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4576..4593)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4747..5335)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4827..4846)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5506..6297,6298..6366))
                     /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     6129..6148
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6367..6471)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttaccggt
      901 caccatggac tacaaagacg atgacgacaa gcttgcggcc gcgaattcag gaaacaaggt
      961 caagaggaca tcctgcatgt atggggcaaa ctgctatagg aagaatcctg ttcattttca
     1021 acattttagc catcctggtg atagtgatta tggaggtgtt caaatcgtgg gccaagatga
     1081 gactgatgac cggcctgaat gtccctatgg accatcctgt tataggaaga atccccagca
     1141 caagatagaa tatagacata atacgcttcc agtgagaaat gttttagatg aatcggtacc
     1201 agtcgactct agaagagcta gcatgactgg tggacagcaa atgggtcggg atctgtacga
     1261 cgatgacgat aaggatccga tggtgtctaa gggcgaagag ctgatcaagg aaaatatgcg
     1321 tatgaaggtg gtcatggaag gttcggtcaa cggccaccaa ttcaaatgca caggtgaagg
     1381 agaaggcaat ccgtacatgg gaactcaaac catgaggatc aaagtcatcg agggaggacc
     1441 cctgccattt gcctttgaca ttcttgccac gtcgttcatg tatggcagcc gtacttttat
     1501 caagtacccg aaaggcattc ctgatttctt taaacagtcc tttcctgagg gttttacttg
     1561 ggaaagagtt acgagatacg aagatggtgg agtcgtcacc gtcatgcagg acaccagcct
     1621 tgaggatggc tgtctcgttt accacgtcca agtcagaggg gtaaactttc cctccaatgg
     1681 tcccgtgatg cagaagaaga ccaagggttg ggagcctaat acagagatga tgtatccagc
     1741 agatggtggt ctgaggggat acactcatat ggcactgaaa gttgatggtg gtggccatct
     1801 gtcttgctct ttcgtaacaa cttacaggtc aaaaaagacc gtcgggaaca tcaagatgcc
     1861 cggtatccat gccgttgatc accgcctgga aaggttagag gaaagtgaca atgaaatgtt
     1921 cgtagtacaa cgcgaacacg cagttgccaa gttcgccggg cttggtggtg ggatggacga
     1981 gctgtacaag taagaattct gcagatatcc atcacactgg cggccgctcg agcatgcatc
     2041 tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg
     2101 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     2161 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     2221 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     2281 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     2341 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     2401 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     2461 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2521 ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     2581 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2641 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     2701 ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa
     2761 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     2821 agggtgtgga aagtccccag gctccccagg caggcagaag tatgcaaagc atgcatctca
     2881 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2941 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3001 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3061 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3121 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     3181 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3241 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3301 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3361 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     3421 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3481 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3541 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3601 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3661 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3721 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     3781 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     3841 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     3901 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     3961 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     4021 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4081 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4141 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     4201 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4261 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     4321 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4381 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4441 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4501 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4561 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4621 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4681 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4741 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4801 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4861 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4921 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4981 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5041 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5101 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5161 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     5221 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5281 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5341 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5401 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5461 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5521 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5581 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5641 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5701 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5761 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5821 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5881 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5941 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6001 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6061 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6121 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6181 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6241 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6301 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6361 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6421 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6481 ccgaaaagtg ccacctgacg tc
//
