LOCUS       sequence-494916-        8468 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression construct for N-terminal S 
            protein-Flag-Streptavidin binding peptide (SFB)-tagged XPF D458-655.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8468)
  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 8468)
  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-D458-655
FEATURES             Location/Qualifiers
     source          1..8468
                     /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(3098..3122)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3305..3322)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     complement(3384..3404)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3389..3718
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3569..3704
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3728..4327
                     /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(3728..3747)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4224..4244
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4548..5017
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5147..5164)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5318..5906)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5398..5417)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6066..6857,6858..6926))
                     /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     6689..6708
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6927..7031)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             7062..7531
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    7596..7795
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             7996..8412
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     8342..8364
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     8380..8396
                     /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 tgaaatttag gaaggaagac ctagtaagag gcacagcatc tgcagatgtt
     2341 tccactgaca ctcggaaagc cggtggccag gaacagaatg gtacacagca aagcatagtt
     2401 gtggatatgc gtgaatttcg aagtgagctt ccatctctga tccatcgtcg gggcattgac
     2461 attgaacccg tgactttaga ggttggagat tacatcctca ctccagaaat gtgcgtggag
     2521 cgcaagagta tcagtgattt aatcggctct ttaaataacg gccgcctcta cagccagtgc
     2581 atctccatgt cccgctacta caagcgtccc gtgcttctga ttgagtttga ccctagcaag
     2641 cctttctctc tcacttcccg aggtgccttg tttcaggaga tctccagcaa tgacattagt
     2701 tccaaactca ctcttcttac acttcacttc cccagactac ggattctctg gtgcccctct
     2761 cctcatgcaa cggcggagtt gtttgaggag ctgaaacaaa gcaagccaca gcctgatgcg
     2821 gcgacagcac tggccattac agcagattct gaaacccttc ccgagtcaga gaagtataat
     2881 cctggtcccc aagacttctt gttaaaaatg ccaggggtga atgccaaaaa ctgccgctcc
     2941 ttgatgcacc acgttaagaa catcgcagaa ttagcagccc tgtcacaaga cgagctcacg
     3001 agtattctgg ggaatgctgc aaatgccaaa cagctttatg atttcattca cacctctttt
     3061 gcagaagtcg tatcaaaagg aaaagggaaa aagtgaaacc cagctttctt gtacaaagtg
     3121 gctcgagcag ggatccgccc ctctccctcc ccccccccta acgttactgg ccgaagccgc
     3181 ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt
     3241 ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt
     3301 tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc gtaggaattc
     3361 gccagcacag tggtcgaccc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3421 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3481 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3541 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3601 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     3661 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     3721 gcttaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag
     3781 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     3841 tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     3901 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     3961 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     4021 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     4081 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg
     4141 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     4201 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     4261 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     4321 tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa aggagcgcac gaccccatgc
     4381 atcgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac
     4441 ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata
     4501 actgagaata gagaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc
     4561 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac
     4621 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     4681 aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat
     4741 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     4801 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
     4861 acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc
     4921 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
     4981 tctgagtgat tgactacccg tcagcggggg tctttcacat gcagcatgta tcaaaattaa
     5041 tttggttttt tttcttaagt atttacatta aatggccata gttgcattaa tgaatcggcc
     5101 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5161 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5221 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5281 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5341 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5401 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5461 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5521 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5581 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5641 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5701 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5761 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5821 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5881 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5941 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6001 tcacctagat ccttttgcgg ccgcaaatca atctaaagta tatatgagta aacttggtct
     6061 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6121 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6181 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6241 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6301 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6361 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6421 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6481 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6541 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6601 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6661 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6721 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6781 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6841 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6901 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6961 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7021 ggggttccgc gcacatttcc ccgaaaagtg ccacctgcag cctgaatatg ggccaaacag
     7081 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga
     7141 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac
     7201 agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc
     7261 agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc
     7321 ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct
     7381 cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa
     7441 ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag
     7501 tgattgacta cccgtcagcg ggggtctttc atttgggggc tcgtccggga tcgggagacc
     7561 cctgcccagg gaccaccgac ccaccaccgg gaggcaagct ggccagcaac ttatctgtgt
     7621 ctgtccgatt gtctagtgtc tatgactgat tttatgcgcc tgcgtcggta ctagttagct
     7681 aactagctct gtatctggcg gacccgtggt ggaactgacg agttctgaac acccggccgc
     7741 aaccctggga gacgtcccag ggactttggg ggccgttttt gtggcccgac ctgaggaagg
     7801 gagtcgatgt ggaatccgac cccgtcagga tatgtggttc tggtaggaga cgagaaccta
     7861 aaacagttcc cgcctccgtc tgaatttttg ctttcggttt ggaaccgaag ccgcgcgtct
     7921 tgtctgctgc agcgctgcag catcgttctg tgttgtctct gtctgactgt gtttctgtat
     7981 ttgtctgaaa attagggcca gactgttacc actcccttaa gtttgacctt aggtcactgg
     8041 aaagatgtcg agcggatcgc tcacaaccag tcggtagatg tcaagaagag acgttgggtt
     8101 accttctgct ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga cggcaccttt
     8161 aaccgagacc tcatcaccca ggttaagatc aaggtctttt cacctggccc gcatggacac
     8221 ccagaccagg tcccctacat cgtgacctgg gaagccttgg cttttgaccc ccctccctgg
     8281 gtcaagccct ttgtacaccc taagcctccg cctcctcttc ctccatccgc cccgtctctc
     8341 ccccttgaac ctcctcgttc gaccccgcct cgatcctccc tttatccagc cctcactcct
     8401 tctctaggcg ccggccggat cccagtgtgg tggtactaat agtaatcaat tacggggtca
     8461 ttagttca
//
