LOCUS       sequence-494940-        9062 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression construct for N-terminal S 
            protein-Flag-Streptavidin binding peptide (SFB)-tagged XPF R454W.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9062)
  AUTHORS   Zhang H, Chen Z, Ye Y, Ye Z, Cao D, Xiong Y, Srivastava M, Feng X, 
            Tang M, Wang C, Tainer JA, Chen J
  TITLE     SLX4IP acts with SLX4 and XPF-ERCC1 to promote interstrand crosslink
            repair.
  JOURNAL   Nucleic Acids Res. 2019 Nov 4;47(19):10181-10201. doi: 
            10.1093/nar/gkz769.
  PUBMED    31495888
REFERENCE   2  (bases 1 to 9062)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2019-11-4- 4"; volume: "47"; issue: 
            "19"; pages: "10181-10201"
COMMENT     Sequence Label: SFB-XPF-R454W
FEATURES             Location/Qualifiers
     source          1..9062
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        22..325
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        326..529
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     480..500
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             589..633
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             667..690
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             694..717
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             727..840
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     protein_bind    913..937
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3692..3716)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3899..3916)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     complement(3978..3998)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3983..4312
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4163..4298
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4322..4921
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4322..4341)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4818..4838
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5142..5611
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5741..5758)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5912..6500)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5992..6011)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6660..7451,7452..7520))
                     /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     7283..7302
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7521..7625)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             7656..8125
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    8190..8389
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             8590..9006
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     8936..8958
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     8974..8990
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
       61 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      121 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      181 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      241 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      301 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      361 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      421 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      481 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
      541 ccgtcagatc cgctagcgct accggactca gatctcgagg ccaccatgaa agaaaccgct
      601 gctgctaaat tcgaacgcca gcacatggac agcggagccg gcgcaggagc cggcgctgac
      661 gcgcctgact acaaagacga tgacgacaag ggagattaca aggatgacga tgacaaaggc
      721 gcgtcgatgg acgagaagac caccggctgg cggggcggcc acgtggtgga gggcctggcc
      781 ggcgagctgg agcagctgcg ggccaggctg gagcaccacc ctcagggcca gcgggagccc
      841 tccggcggct gcaagctggg cggcgcgcct gaattctgca gtcgacggta ccgcgggccc
      901 atcgctcgag agacaagttt gtacaaaaaa gcaggcttca tggagtcagg gcagccggct
      961 cgacggattg ccatggcgcc gctgctggag tacgagcgac agctggtgct ggaactgctc
     1021 gacactgacg ggctagtagt gtgcgcccgc gggctcggcg cggaccggct cctctaccac
     1081 tttctccagc tgcactgcca cccagcctgc ctggtgctgg tgctcaacac gcagccggcc
     1141 gaggaggagt attttatcaa tcagctgaag atagaaggag ttgaacacct ccctcgccgt
     1201 gtaacaaatg aaatcacaag caacagtcgc tatgaagttt acacacaagg tggtgttata
     1261 tttgcgacaa gtaggatact tgtggttgac ttcttgactg atagaatacc ttcagattta
     1321 attactggca tcttggtgta tagagcccac agaataatcg agtcttgtca agaagcattc
     1381 atcttgcgcc tctttcgcca gaaaaacaaa cgtggtttta ttaaagcttt cacagacaat
     1441 gctgttgcct ttgatactgg tttttgtcat gtggaaagag tgatgagaaa tctttttgtg
     1501 aggaaactgt atctgtggcc aaggttccat gtagcagtaa actcattttt agaacagcac
     1561 aaacctgaag ttgtagaaat ccatgtttct atgacaccta ccatgcttgc tatacagact
     1621 gctatactgg acattttaaa tgcatgtcta aaggaactaa aatgccataa cccatcgctt
     1681 gaagtggaag atttatcttt agaaaatgct attggaaaac cttttgacaa gacaatccgc
     1741 cattatctgg atcctttgtg gcaccagctt ggagccaaga ctaaatcctt agttcaggat
     1801 ttgaagatat tacgaacttt gctgcagtat ctctctcagt atgattgtgt cacatttctt
     1861 aatcttctgg aatctctgag agcaacggaa aaagcttttg gtcagaattc aggttggctg
     1921 tttcttgact ccagcacctc gatgtttata aatgctcgag caagggttta tcatcttcca
     1981 gatgccaaaa tgagtaaaaa agaaaaaata tctgaaaaaa tggaaattaa agaaggggaa
     2041 gaaacaaaaa aggaactggt cctagaaagc aacccaaagt gggaggcact gactgaagta
     2101 ttaaaagaaa ttgaggcaga aaataaggag agtgaagctc ttggtggtcc aggtcaagta
     2161 ctgatttgtg caagtgatga ccgaacatgt tcccagctga gagactatat cactcttgga
     2221 gcggaggcct tcttattgag gctctacagg aaaacctttg agaaggatag caaagctgaa
     2281 gaagtctgga tgaaattttg gaaggaagac agttcaaaga gaattaggaa atctcacaaa
     2341 agacctaaag acccccaaaa caaagaacgg gcttctacca aagaaagaac cctcaaaaag
     2401 aaaaaacgga agttgacctt aactcaaatg gtaggaaaac ctgaagaact ggaagaggaa
     2461 ggagatgtcg aggaaggata tcgtcgagaa ataagcagta gcccagaaag ctgcccggaa
     2521 gaaattaagc atgaagaatt tgatgtaaat ttgtcatcgg atgctgcttt cggaatcctg
     2581 aaagaacccc tcactatcat ccatccgctt ctgggttgca gcgaccccta tgctctgaca
     2641 agggtactac atgaagtgga gccaagatac gtggttcttt atgacgcaga gctaaccttt
     2701 gttcggcagc ttgaaattta cagggcgagt aggcctggga aacctctgag ggtttacttt
     2761 cttatatacg gaggttcaac tgaggaacaa cgctatctca ctgctttgcg gaaagaaaag
     2821 gaagcttttg aaaaactcat aagggaaaaa gcaagcatgg ttgtccctga agaaagagaa
     2881 ggcagagatg aaacaaactt agacctagta agaggcacag catctgcaga tgtttccact
     2941 gacactcgga aagccggtgg ccaggaacag aatggtacac agcaaagcat agttgtggat
     3001 atgcgtgaat ttcgaagtga gcttccatct ctgatccatc gtcggggcat tgacattgaa
     3061 cccgtgactt tagaggttgg agattacatc ctcactccag aaatgtgcgt ggagcgcaag
     3121 agtatcagtg atttaatcgg ctctttaaat aacggccgcc tctacagcca gtgcatctcc
     3181 atgtcccgct actacaagcg tcccgtgctt ctgattgagt ttgaccctag caagcctttc
     3241 tctctcactt cccgaggtgc cttgtttcag gagatctcca gcaatgacat tagttccaaa
     3301 ctcactcttc ttacacttca cttccccaga ctacggattc tctggtgccc ctctcctcat
     3361 gcaacggcgg agttgtttga ggagctgaaa caaagcaagc cacagcctga tgcggcgaca
     3421 gcactggcca ttacagcaga ttctgaaacc cttcccgagt cagagaagta taatcctggt
     3481 ccccaagact tcttgttaaa aatgccaggg gtgaatgcca aaaactgccg ctccttgatg
     3541 caccacgtta agaacatcgc agaattagca gccctgtcac aagacgagct cacgagtatt
     3601 ctggggaatg ctgcaaatgc caaacagctt tatgatttca ttcacacctc ttttgcagaa
     3661 gtcgtatcaa aaggaaaagg gaaaaagtga aacccagctt tcttgtacaa agtggctcga
     3721 gcagggatcc gcccctctcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa
     3781 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat
     3841 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct
     3901 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcgtagga attcgccagc
     3961 acagtggtcg accctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4021 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4081 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4141 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4201 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     4261 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttac
     4321 catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
     4381 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
     4441 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     4501 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     4561 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     4621 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     4681 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     4741 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     4801 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     4861 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     4921 acgcccgccc cacgacccgc agcgcccgac cgaaaggagc gcacgacccc atgcatcgat
     4981 aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag
     5041 gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata cataactgag
     5101 aatagagaag ttcagatcaa ggtcaggaac agatggaaca gctgaatatg ggccaaacag
     5161 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga
     5221 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac
     5281 agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc
     5341 agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc
     5401 ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct
     5461 cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa
     5521 ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag
     5581 tgattgacta cccgtcagcg ggggtctttc acatgcagca tgtatcaaaa ttaatttggt
     5641 tttttttctt aagtatttac attaaatggc catagttgca ttaatgaatc ggccaacgcg
     5701 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5761 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5821 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5881 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5941 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6001 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6061 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6121 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6181 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6241 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6301 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6361 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6421 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6481 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6541 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6601 agatcctttt gcggccgcaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6661 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6721 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6781 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6841 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6901 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6961 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7021 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7081 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7141 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7201 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7261 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7321 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7381 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7441 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7501 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7561 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7621 ccgcgcacat ttccccgaaa agtgccacct gcagcctgaa tatgggccaa acaggatatc
     7681 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag ctgaatatgg
     7741 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
     7801 tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt ttccagggtg
     7861 ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg
     7921 cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg
     7981 gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa taaaccctct
     8041 tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg
     8101 actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga gacccctgcc
     8161 cagggaccac cgacccacca ccgggaggca agctggccag caacttatct gtgtctgtcc
     8221 gattgtctag tgtctatgac tgattttatg cgcctgcgtc ggtactagtt agctaactag
     8281 ctctgtatct ggcggacccg tggtggaact gacgagttct gaacacccgg ccgcaaccct
     8341 gggagacgtc ccagggactt tgggggccgt ttttgtggcc cgacctgagg aagggagtcg
     8401 atgtggaatc cgaccccgtc aggatatgtg gttctggtag gagacgagaa cctaaaacag
     8461 ttcccgcctc cgtctgaatt tttgctttcg gtttggaacc gaagccgcgc gtcttgtctg
     8521 ctgcagcgct gcagcatcgt tctgtgttgt ctctgtctga ctgtgtttct gtatttgtct
     8581 gaaaattagg gccagactgt taccactccc ttaagtttga ccttaggtca ctggaaagat
     8641 gtcgagcgga tcgctcacaa ccagtcggta gatgtcaaga agagacgttg ggttaccttc
     8701 tgctctgcag aatggccaac ctttaacgtc ggatggccgc gagacggcac ctttaaccga
     8761 gacctcatca cccaggttaa gatcaaggtc ttttcacctg gcccgcatgg acacccagac
     8821 caggtcccct acatcgtgac ctgggaagcc ttggcttttg acccccctcc ctgggtcaag
     8881 ccctttgtac accctaagcc tccgcctcct cttcctccat ccgccccgtc tctccccctt
     8941 gaacctcctc gttcgacccc gcctcgatcc tccctttatc cagccctcac tccttctcta
     9001 ggcgccggcc ggatcccagt gtggtggtac taatagtaat caattacggg gtcattagtt
     9061 ca
//
