LOCUS       sequence-450092-        6936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminally mEGFP-tagged PARP11.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6936)
  AUTHORS   Weixler L, Ikenga NJ, Voorneveld J, Aydin G, Bolte TM, Momoh J, 
            Butepage M, Golzmann A, Luscher B, Filippov DV, Zaja R, Feijs KL
  TITLE     Protein and RNA ADP-ribosylation detection is influenced by sample 
            preparation and reagents used.
  JOURNAL   Life Sci Alliance. 2022 Nov 11;6(1):e202201455. doi: 
            10.26508/lsa.202201455.
  PUBMED    36368907
REFERENCE   2  (bases 1 to 6936)
  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: "Life Sci 
            Alliance."; date: "2022-11-11"; volume: "6(1):e202201455. doi"; 
            pages: " 10.26508/lsa.202201455"
COMMENT     Sequence Label: pcDNA5-FRT/TO-N-mEGFP-PARP11
FEATURES             Location/Qualifiers
     source          1..6936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    7..31
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1052..1076)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1190..1207)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1196..1420
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1553..1572)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    1704..1751
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             1759..2781
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             1759..2781
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    2911..3032
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2948..2967)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3002..3021
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3081..3097)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3081..3097)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3094..3116)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3105..3121
                     /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(3129..3135,3136..3153,3154..3159))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3174..3195
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3312..3329)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3483..4071)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3563..3582)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4242..5033,5034..5102))
                     /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     4865..4884
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5103..5207)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5282..5301)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5473..5852
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5853..6056
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6007..6027
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    6058..6076
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6079..6097
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     6107..6131
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      6214..6223
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6220..6222,6223..6225,6226..6936)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6265..6286)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6526..6545)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6873..6894
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gctatcacaa gtttgtacaa aaaagcaggc ttcatgtggg aagcgaatcc agagatgttt
       61 cacaaagcag aagaattatt ttctaaaaca acaaacaatg aagtggatga catggacacg
      121 tcagataccc agtggggctg gttttacttg gcagaatgtg ggaagtggca catgtttcag
      181 ccggatacca acagtcagtg ttcagttagc agtgaagata tcgaaaaaag cttcaaaaca
      241 aacccttgtg gctccatttc ttttactact tccaaattca gctacaagat agactttgca
      301 gaaatgaagc aaatgaatct caccactgga aagcagcgct taataaaaag agcccccttt
      361 tctatcagtg ctttcagtta catctgtgaa aacgaggcca tccctatgcc accacactgg
      421 gagaatgtga atactcaagt accatatcag cttattcctc tgcacaatca aacacatgaa
      481 tataatgaag ttgctaatct ctttgggaag acgatggatc gcaaccgaat taaaagaatt
      541 cagagaattc aaaacctaga tttgtgggag ttcttttgca ggaaaaaggc tcagctcaag
      601 aaaaaaagag gtgtgcctca gattaatgaa caaatgctgt ttcatggtac cagcagtgaa
      661 tttgtggaag caatctgcat tcataacttt gattggagaa taaatggtat acatggtgct
      721 gtctttggaa aaggaaccta ttttgctaga gatgctgctt attccagtcg tttctgcaaa
      781 gatgacataa agcatgggaa cacattccaa attcatggtg tcagcttgca acagcggcat
      841 ctgtttagaa catataaatc tatgtttctt gctcgagtgc taattggaga ttacataaac
      901 ggagactcca aatacatgcg acctccttcc aaagacggga gctatgtgaa tttatatgac
      961 agctgtgtgg atgatacctg gaacccaaag atctttgtgg tttttgatgc caaccaaatc
     1021 tatcctgagt acttgataga ctttcattga tacccagctt tcttgtacaa agtggtgata
     1081 gcttggtacc gagctcggat ccactagtcc agtgtggtgg aattctgcag atatccagca
     1141 cagtggcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg
     1201 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     1261 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     1321 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     1381 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     1441 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     1501 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     1561 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     1621 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     1681 tagggtgatg gttcacgtac ctagaagttc ctattccgaa gttcctattc tctagaaagt
     1741 ataggaactt ccttggccaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg
     1801 atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt
     1861 gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat
     1921 ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg
     1981 gaagtgcttg acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca
     2041 cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc
     2101 gcggaggcca tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca
     2161 ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct
     2221 gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg
     2281 caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg
     2341 cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt
     2401 gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg
     2461 aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag
     2521 cttgcaggat cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat
     2581 cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca
     2641 atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc
     2701 gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact
     2761 cgtccgaggg caaaggaata gcacgtacta cgagatttcg attccaccgc cgccttctat
     2821 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     2881 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     2941 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3001 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     3061 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     3121 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     3181 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     3241 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3301 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3361 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3421 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3481 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3541 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3601 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3661 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3721 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3781 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3841 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3901 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     3961 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4021 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4081 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4141 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4201 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4261 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4321 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4381 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4441 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4501 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     4561 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     4621 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     4681 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     4741 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     4801 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     4861 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     4921 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     4981 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5041 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5101 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5161 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5221 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca
     5281 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
     5341 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
     5401 attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca
     5461 gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
     5521 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     5581 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     5641 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     5701 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     5761 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     5821 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
     5881 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     5941 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
     6001 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctcc
     6061 ctatcagtga tagagatctc cctatcagtg atagagatcg tcgacgagct cgtttagtga
     6121 accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg
     6181 accgatccag cctccggact ctagcgttta aacgccacca tggtgagcaa gggcgaggag
     6241 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     6301 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     6361 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     6421 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     6481 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     6541 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     6601 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     6661 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     6721 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     6781 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccaag
     6841 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     6901 gccgggatca ctctcggcat ggacgagctg tacaag
//
