LOCUS       sequence-494919-        8588 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression construct for N-terminal S 
            protein-Flag-Streptavidin binding peptide (SFB)-tagged XPF D656-813.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8588)
  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 8588)
  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-D656-813
FEATURES             Location/Qualifiers
     source          1..8588
                     /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(3218..3242)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3425..3442)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     complement(3504..3524)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3509..3838
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3689..3824
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3848..4447
                     /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(3848..3867)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4344..4364
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4668..5137
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5267..5284)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5438..6026)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5518..5537)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6186..6977,6978..7046))
                     /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     6809..6828
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7047..7151)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             7182..7651
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    7716..7915
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             8116..8532
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     8462..8484
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     8500..8516
                     /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 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 agacgaggag ctgaaacaaa gcaagccaca gcctgatgcg
     2941 gcgacagcac tggccattac agcagattct gaaacccttc ccgagtcaga gaagtataat
     3001 cctggtcccc aagacttctt gttaaaaatg ccaggggtga atgccaaaaa ctgccgctcc
     3061 ttgatgcacc acgttaagaa catcgcagaa ttagcagccc tgtcacaaga cgagctcacg
     3121 agtattctgg ggaatgctgc aaatgccaaa cagctttatg atttcattca cacctctttt
     3181 gcagaagtcg tatcaaaagg aaaagggaaa aagtgaaacc cagctttctt gtacaaagtg
     3241 gctcgagcag ggatccgccc ctctccctcc ccccccccta acgttactgg ccgaagccgc
     3301 ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt
     3361 ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt
     3421 tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc gtaggaattc
     3481 gccagcacag tggtcgaccc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3541 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3601 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3661 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3721 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     3781 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     3841 gcttaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag
     3901 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     3961 tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     4021 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     4081 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     4141 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     4201 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg
     4261 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     4321 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     4381 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     4441 tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa aggagcgcac gaccccatgc
     4501 atcgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac
     4561 ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata
     4621 actgagaata gagaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc
     4681 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac
     4741 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     4801 aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat
     4861 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     4921 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
     4981 acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc
     5041 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
     5101 tctgagtgat tgactacccg tcagcggggg tctttcacat gcagcatgta tcaaaattaa
     5161 tttggttttt tttcttaagt atttacatta aatggccata gttgcattaa tgaatcggcc
     5221 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5281 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5341 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5401 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5461 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5521 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5581 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5641 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5701 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5761 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5821 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5881 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5941 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6001 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6061 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6121 tcacctagat ccttttgcgg ccgcaaatca atctaaagta tatatgagta aacttggtct
     6181 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6241 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6301 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6361 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6421 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6481 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6541 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6601 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6661 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6721 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6781 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6841 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6901 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6961 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7021 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7081 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7141 ggggttccgc gcacatttcc ccgaaaagtg ccacctgcag cctgaatatg ggccaaacag
     7201 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga
     7261 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac
     7321 agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc
     7381 agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc
     7441 ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct
     7501 cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa
     7561 ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag
     7621 tgattgacta cccgtcagcg ggggtctttc atttgggggc tcgtccggga tcgggagacc
     7681 cctgcccagg gaccaccgac ccaccaccgg gaggcaagct ggccagcaac ttatctgtgt
     7741 ctgtccgatt gtctagtgtc tatgactgat tttatgcgcc tgcgtcggta ctagttagct
     7801 aactagctct gtatctggcg gacccgtggt ggaactgacg agttctgaac acccggccgc
     7861 aaccctggga gacgtcccag ggactttggg ggccgttttt gtggcccgac ctgaggaagg
     7921 gagtcgatgt ggaatccgac cccgtcagga tatgtggttc tggtaggaga cgagaaccta
     7981 aaacagttcc cgcctccgtc tgaatttttg ctttcggttt ggaaccgaag ccgcgcgtct
     8041 tgtctgctgc agcgctgcag catcgttctg tgttgtctct gtctgactgt gtttctgtat
     8101 ttgtctgaaa attagggcca gactgttacc actcccttaa gtttgacctt aggtcactgg
     8161 aaagatgtcg agcggatcgc tcacaaccag tcggtagatg tcaagaagag acgttgggtt
     8221 accttctgct ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga cggcaccttt
     8281 aaccgagacc tcatcaccca ggttaagatc aaggtctttt cacctggccc gcatggacac
     8341 ccagaccagg tcccctacat cgtgacctgg gaagccttgg cttttgaccc ccctccctgg
     8401 gtcaagccct ttgtacaccc taagcctccg cctcctcttc ctccatccgc cccgtctctc
     8461 ccccttgaac ctcctcgttc gaccccgcct cgatcctccc tttatccagc cctcactcct
     8521 tctctaggcg ccggccggat cccagtgtgg tggtactaat agtaatcaat tacggggtca
     8581 ttagttca
//
