LOCUS       sequence-450104-        7893 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminally mEGFP-tagged PARP7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7893)
  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 7893)
  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-PARP7
FEATURES             Location/Qualifiers
     source          1..7893
                     /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(2009..2033)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2147..2164)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2153..2377
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2510..2529)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    2661..2708
                     /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             2716..3738
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             2716..3738
                     /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    3868..3989
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3905..3924)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3959..3978
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4038..4054)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4038..4054)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4051..4073)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4062..4078
                     /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(4086..4092,4093..4110,4111..4116))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4131..4152
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4269..4286)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4440..5028)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4520..4539)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5199..5990,5991..6059))
                     /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     5822..5841
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6060..6164)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6239..6258)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6430..6809
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6810..7013
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6964..6984
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    7015..7033
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7036..7054
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     7064..7088
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      7171..7180
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(7177..7179,7180..7182,7183..7893)
                     /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(7222..7243)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7483..7502)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7830..7851
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gctatcacaa gtttgtacaa aaaagcaggc ttcatggaaa tggaaaccac cgaacctgag
       61 ccagactgtg tagtgcagcc tccctctcct cctgatgact tttcatgcca aatgagactc
      121 tctgagaaga tcactccatt gaagacttgt tttaagaaaa aggatcagaa aagattggga
      181 actggaaccc tgaggtcttt gaggccaata ttaaacactc ttctagaatc tggctcactt
      241 gatggggttt ttagatctag gaaccagagt acagatgaga acagcttaca tgaacctatg
      301 atgaagaaag ccatggaaat caattcatca tgcccaccag cagaaaataa tatgtctgtt
      361 ctgattcctg ataggacaaa tgttggggac cagataccgg aagcccatcc ttccactgaa
      421 gctccagaac gagtggttcc aatccaagat cacagctttc catcagaaac cctcagtggg
      481 acggtggcag attccacacc agctcacttc cagactgatc ttttgcaccc agtttcaagt
      541 gatgttccta ctagtcctga ctgcttagac aaagtcatag attatgttcc aggcattttc
      601 caagaaaaca gttttacaat ccaatacatt ctggacacca gtgataagct gagtactgag
      661 ctctttcagg acaaaagtga agaggcttcc cttgacctcg tgtttgagct ggtgaaccag
      721 ttgcagtacc acactcacca agagaacgga attgaaattt gcatggactt tctgcaaggc
      781 acttgtattt atggcaggga ttgtttgaag caccacactg tcttgccata tcattggcag
      841 atcaaaagga caactactca aaagtggcag agtgtattca atgattctca ggagcacttg
      901 gaaagatttt actgtaaccc agaaaatgat agaatgagaa tgaagtatgg aggacaagaa
      961 ttttgggcag atttgaatgc catgaacgtg tatgaaacaa ctgaatttga ccaactacga
     1021 aggctgtcca caccaccctc tagcaatgtc aactctattt accacacagt ctggaaattc
     1081 ttctgtaggg accactttgg atggagagag tatcccgagt ctgtcattcg attgattgaa
     1141 gaagccaact ctcggggtct gaaagaggtt cgatttatga tgtggaataa ccactacatc
     1201 ctccacaatt cattcttcag gagagagata aaaaggagac ccctcttccg ctcctgtttt
     1261 atactgcttc catatttaca gacacttggt ggggttccca cacaagctcc tccacctctt
     1321 gaagcaactt catcatcaca aattatctgc ccagatgggg tcacttcagc aaacttttac
     1381 cctgaaactt gggtttatat gcatccatct caggacttca tccaagtccc tgtttctgca
     1441 gaggataaaa gttatcggat catttacaat ctttttcata agactgtgcc tgagtttaaa
     1501 tacagaattt tgcagatatt gagagtccaa aaccagtttc tttgggagaa atataaaagg
     1561 aaaaaggaat atatgaacag gaaaatgttt ggccgtgaca ggataataaa tgagagacat
     1621 ttatttcatg gaacatccca ggatgtggta gatggaatct gcaaacacaa ctttgaccct
     1681 cgagtctgtg gaaagcatgc tacaatgttt ggacaaggca gttattttgc aaagaaggca
     1741 agctactctc ataacttttc taagaagtcc tccaaaggag tccacttcat gtttctggcc
     1801 aaagtgctga cgggcagata cacaatgggc agtcatggca tgagaaggcc cccgccagtc
     1861 aatcctggca gtgtcaccag tgacctttat gactcttgtg tggataattt ctttgagcct
     1921 cagatttttg tcatttttaa tgatgaccag agttaccctt attttgttat ccaatatgaa
     1981 gaagtcagta acactgtttc catttgatac ccagctttct tgtacaaagt ggtgatagct
     2041 tggtaccgag ctcggatcca ctagtccagt gtggtggaat tctgcagata tccagcacag
     2101 tggcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct
     2161 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     2221 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2281 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     2341 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     2401 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2461 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2521 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2581 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     2641 ggtgatggtt cacgtaccta gaagttccta ttccgaagtt cctattctct agaaagtata
     2701 ggaacttcct tggccaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc
     2761 gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct
     2821 ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt
     2881 ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa
     2941 gtgcttgaca ttggggaatt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag
     3001 ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg
     3061 gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc
     3121 ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat
     3181 ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag
     3241 gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac
     3301 gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac
     3361 tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg
     3421 ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt
     3481 gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag
     3541 agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc
     3601 gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc
     3661 tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt
     3721 ccgagggcaa aggaatagca cgtactacga gatttcgatt ccaccgccgc cttctatgaa
     3781 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     3841 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     3901 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     3961 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     4021 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4081 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4141 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4201 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4261 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4321 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4381 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4441 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4501 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4561 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4621 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4681 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4741 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4801 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4861 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4921 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4981 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5041 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5101 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5161 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5221 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5281 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5341 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5401 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5461 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5521 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5581 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5641 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5701 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5761 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     5821 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     5881 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     5941 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6001 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6061 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6121 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6181 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta
     6241 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     6301 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     6361 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat
     6421 atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     6481 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     6541 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     6601 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     6661 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     6721 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     6781 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     6841 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     6901 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     6961 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctcccta
     7021 tcagtgatag agatctccct atcagtgata gagatcgtcg acgagctcgt ttagtgaacc
     7081 gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc
     7141 gatccagcct ccggactcta gcgtttaaac gccaccatgg tgagcaaggg cgaggagctg
     7201 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     7261 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     7321 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     7381 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     7441 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     7501 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     7561 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     7621 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     7681 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     7741 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccaagctg
     7801 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     7861 gggatcactc tcggcatgga cgagctgtac aag
//
