LOCUS       sequence-450105-        7815 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminally mEGFP-tagged PARP6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7815)
  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 7815)
  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-PARP6
FEATURES             Location/Qualifiers
     source          1..7815
                     /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(1931..1955)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2069..2086)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2075..2299
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2432..2451)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    2583..2630
                     /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             2638..3660
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             2638..3660
                     /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    3790..3911
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3827..3846)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3881..3900
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3960..3976)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3960..3976)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3973..3995)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3984..4000
                     /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(4008..4014,4015..4032,4033..4038))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4053..4074
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4191..4208)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4362..4950)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4442..4461)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5121..5912,5913..5981))
                     /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     5744..5763
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5982..6086)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6161..6180)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6352..6731
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6732..6935
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6886..6906
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    6937..6955
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6958..6976
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     6986..7010
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      7093..7102
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(7099..7101,7102..7104,7105..7815)
                     /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(7144..7165)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7405..7424)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7752..7773
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gctatcacaa gtttgtacaa aaaagcaggc ttcatggaca tcaaaggcca gttctggaat
       61 gatgacgact cggagggaga taatgaatca gaggaatttc tctatggcgt tcaggggagc
      121 tgtgcagctg acctgtatcg acacccacag cttgatgcag acattgaagc cgtgaaggag
      181 atctacagtg agaactctgt atccatcaga gaatatggaa ctatcgatga cgtggacatt
      241 gacctccaca tcaacatcag cttcctcgat gaggaagtct ctacagcctg gaaggtcctc
      301 cggacagaac ctattgtgtt gaggctgcga ttttctctct cccagtacct agatggacca
      361 gaaccatcca ttgaggtttt ccagccatca aataaggaag gatttgggct gggtcttcag
      421 ttgaaaaaga tcctgggtat gtttacatcc caacaatgga aacatctgag caatgatttc
      481 ttgaagaccc agcaggagaa gaggcacagt tggttcaagg caagtggtac catcaagaag
      541 ttccgagctg gcctcagcat cttttcaccc atccccaagt ctcccagttt ccctatcata
      601 caggactcca tgctgaaagg caaactaggt gtaccagagc ttcgggttgg gcgcctcatg
      661 aaccgttcca tctcctgtac catgaagaac cccaaagtgg aagtgtttgg ctaccctccc
      721 agcccccagg caggtctcct gtgccctcag cacgtgggcc tccctccccc agcacggacc
      781 tctcctttgg tcagtggtca ctgcaagaac attcccactc tggagtatgg attcctcgtt
      841 cagatcatga agtatgcaga acagaggatt ccaacattga atgagtactg tgtggtgtgt
      901 gatgagcagc atgtcttcca aaatggatct atgctgaagc cagctgtctg tactcgtgaa
      961 ctatgcgttt tctccttcta cacactgggc gtcatgtctg gagctgcaga ggaggtggcc
     1021 actggagcag aggtggtgga tctgctggtg gccatgtgta gggcagcttt agagtcccct
     1081 agaaagagca tcatctttga gccttatccc tctgtggtgg accccactga tcccaagact
     1141 ctggccttta accctaagaa gaagaattat gagcggcttc agaaagctct ggatagtgtg
     1201 atgtctattc gggagatgac ccagggctca tatttggaaa tcaagaaaca gatggacaag
     1261 ttggatcccc tggcccatcc tctcctgcag tggatcatct ctagcaacag gtcacacatt
     1321 gtcaaactac ctctcagcag gcagctgaag ttcatgcaca cctcacacca gttcctcctg
     1381 ctgagcagcc ctcctgccaa ggaggctcgg ttccggaccg ccaagaagct ctatggcagc
     1441 acctttgcct tccatgggtc ccacattgag aactggcatt cgatcctgcg caatgggctg
     1501 gtcaatgcat cctacaccaa actgcagctg catggagcag cctatggcaa aggcatctac
     1561 ctgagcccca tctccagtat ttcctttgga tactcaggaa tgggaaaagg acagcacagg
     1621 atgccctcca aggatgagct ggtccagaga tacaacagga tgaataccat cccccagacc
     1681 cgatccattc agtcacggtt cctgcagagt cggaatctaa actgtatagc actttgtgaa
     1741 gtgattacat ctaaggacct ccagaagcat gggaacatct gggtgtgccc tgtgtccgac
     1801 catgtctgca caagattctt ctttgtatat gaggatggtc aggtgggcga tgccaacatt
     1861 aatactcagg accccaagat acagaaggaa atcatgcgtg tgatcggaac tcaggtttac
     1921 acaaactgat acccagcttt cttgtacaaa gtggtgatag cttggtaccg agctcggatc
     1981 cactagtcca gtgtggtgga attctgcaga tatccagcac agtggcggcc gctcgagtct
     2041 agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     2101 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     2161 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     2221 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     2281 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     2341 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2401 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2461 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2521 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtacc
     2581 tagaagttcc tattccgaag ttcctattct ctagaaagta taggaacttc cttggccaaa
     2641 aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc
     2701 tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga
     2761 gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat
     2821 gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggaa
     2881 ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac
     2941 ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggccat ggatgcgatc
     3001 gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt
     3061 caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg
     3121 caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg
     3181 ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac
     3241 aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc
     3301 ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg
     3361 gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc gccgcggctc
     3421 cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat
     3481 ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg
     3541 actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta
     3601 gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaaggaatag
     3661 cacgtactac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     3721 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     3781 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3841 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3901 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     3961 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4021 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4081 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4141 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4201 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4261 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4321 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4381 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4441 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4501 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4561 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4621 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4681 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4741 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4801 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4861 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4921 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4981 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5041 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5101 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5161 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5221 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5281 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5341 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5401 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5461 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5521 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5581 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5641 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5701 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5761 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5821 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5881 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5941 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6001 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6061 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     6121 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
     6181 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     6241 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     6301 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
     6361 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     6421 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     6481 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     6541 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     6601 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     6661 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     6721 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     6781 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     6841 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     6901 ggcgtgtacg gtgggaggtc tatataagca gagctctccc tatcagtgat agagatctcc
     6961 ctatcagtga tagagatcgt cgacgagctc gtttagtgaa ccgtcagatc gcctggagac
     7021 gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc ctccggactc
     7081 tagcgtttaa acgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     7141 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     7201 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     7261 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     7321 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     7381 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     7441 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     7501 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     7561 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     7621 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     7681 ctgctgcccg acaaccacta cctgagcacc cagtccaagc tgagcaaaga ccccaacgag
     7741 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     7801 gacgagctgt acaag
//
