LOCUS       sequence-494932-        8453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression construct for N-terminal S 
            protein-Flag-Streptavidin binding peptide (SFB)-tagged XPF D1-204.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8453)
  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 8453)
  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-D1-204
FEATURES             Location/Qualifiers
     source          1..8453
                     /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(3083..3107)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3290..3307)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     complement(3369..3389)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3374..3703
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3554..3689
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3713..4312
                     /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(3713..3732)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4209..4229
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4533..5002
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5132..5149)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5303..5891)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5383..5402)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6051..6842,6843..6911))
                     /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     6674..6693
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6912..7016)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             7047..7516
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    7581..7780
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             7981..8397
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     8327..8349
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     8365..8381
                     /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 tggaacagca caaacctgaa
      961 gttgtagaaa tccatgtttc tatgacacct accatgcttg ctatacagac tgctatactg
     1021 gacattttaa atgcatgtct aaaggaacta aaatgccata acccatcgct tgaagtggaa
     1081 gatttatctt tagaaaatgc tattggaaaa ccttttgaca agacaatccg ccattatctg
     1141 gatcctttgt ggcaccagct tggagccaag actaaatcct tagttcagga tttgaagata
     1201 ttacgaactt tgctgcagta tctctctcag tatgattgtg tcacatttct taatcttctg
     1261 gaatctctga gagcaacgga aaaagctttt ggtcagaatt caggttggct gtttcttgac
     1321 tccagcacct cgatgtttat aaatgctcga gcaagggttt atcatcttcc agatgccaaa
     1381 atgagtaaaa aagaaaaaat atctgaaaaa atggaaatta aagaagggga agaaacaaaa
     1441 aaggaactgg tcctagaaag caacccaaag tgggaggcac tgactgaagt attaaaagaa
     1501 attgaggcag aaaataagga gagtgaagct cttggtggtc caggtcaagt actgatttgt
     1561 gcaagtgatg accgaacatg ttcccagctg agagactata tcactcttgg agcggaggcc
     1621 ttcttattga ggctctacag gaaaaccttt gagaaggata gcaaagctga agaagtctgg
     1681 atgaaattta ggaaggaaga cagttcaaag agaattagga aatctcacaa aagacctaaa
     1741 gacccccaaa acaaagaacg ggcttctacc aaagaaagaa ccctcaaaaa gaaaaaacgg
     1801 aagttgacct taactcaaat ggtaggaaaa cctgaagaac tggaagagga aggagatgtc
     1861 gaggaaggat atcgtcgaga aataagcagt agcccagaaa gctgcccgga agaaattaag
     1921 catgaagaat ttgatgtaaa tttgtcatcg gatgctgctt tcggaatcct gaaagaaccc
     1981 ctcactatca tccatccgct tctgggttgc agcgacccct atgctctgac aagggtacta
     2041 catgaagtgg agccaagata cgtggttctt tatgacgcag agctaacctt tgttcggcag
     2101 cttgaaattt acagggcgag taggcctggg aaacctctga gggtttactt tcttatatac
     2161 ggaggttcaa ctgaggaaca acgctatctc actgctttgc ggaaagaaaa ggaagctttt
     2221 gaaaaactca taagggaaaa agcaagcatg gttgtccctg aagaaagaga aggcagagat
     2281 gaaacaaact tagacctagt aagaggcaca gcatctgcag atgtttccac tgacactcgg
     2341 aaagccggtg gccaggaaca gaatggtaca cagcaaagca tagttgtgga tatgcgtgaa
     2401 tttcgaagtg agcttccatc tctgatccat cgtcggggca ttgacattga acccgtgact
     2461 ttagaggttg gagattacat cctcactcca gaaatgtgcg tggagcgcaa gagtatcagt
     2521 gatttaatcg gctctttaaa taacggccgc ctctacagcc agtgcatctc catgtcccgc
     2581 tactacaagc gtcccgtgct tctgattgag tttgacccta gcaagccttt ctctctcact
     2641 tcccgaggtg ccttgtttca ggagatctcc agcaatgaca ttagttccaa actcactctt
     2701 cttacacttc acttccccag actacggatt ctctggtgcc cctctcctca tgcaacggcg
     2761 gagttgtttg aggagctgaa acaaagcaag ccacagcctg atgcggcgac agcactggcc
     2821 attacagcag attctgaaac ccttcccgag tcagagaagt ataatcctgg tccccaagac
     2881 ttcttgttaa aaatgccagg ggtgaatgcc aaaaactgcc gctccttgat gcaccacgtt
     2941 aagaacatcg cagaattagc agccctgtca caagacgagc tcacgagtat tctggggaat
     3001 gctgcaaatg ccaaacagct ttatgatttc attcacacct cttttgcaga agtcgtatca
     3061 aaaggaaaag ggaaaaagtg aaacccagct ttcttgtaca aagtggctcg agcagggatc
     3121 cgcccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg
     3181 tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc
     3241 cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa
     3301 ggaatgcaag gtctgttgaa tgtcgtgaag gaagcgtagg aattcgccag cacagtggtc
     3361 gaccctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3421 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3481 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3541 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3601 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     3661 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctta ccatgaccga
     3721 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
     3781 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
     3841 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     3901 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     3961 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     4021 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     4081 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     4141 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     4201 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     4261 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacgcccgcc
     4321 ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catgcatcga taaaataaaa
     4381 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa
     4441 gctagcttaa gtaacgccat tttgcaaggc atggaaaaat acataactga gaatagagaa
     4501 gttcagatca aggtcaggaa cagatggaac agctgaatat gggccaaaca ggatatctgt
     4561 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggaacagctg aatatgggcc
     4621 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc
     4681 ccagatgcgg tccagccctc agcagtttct agagaaccat cagatgtttc cagggtgccc
     4741 caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt
     4801 ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcgggg
     4861 cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc
     4921 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
     4981 acccgtcagc gggggtcttt cacatgcagc atgtatcaaa attaatttgg ttttttttct
     5041 taagtattta cattaaatgg ccatagttgc attaatgaat cggccaacgc gcggggagag
     5101 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5161 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5221 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5281 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5341 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5401 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5461 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5521 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5581 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5641 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5701 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5761 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5821 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5881 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5941 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6001 tgcggccgca aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6061 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6121 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6181 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6241 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6301 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6361 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6421 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6481 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6541 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6601 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6661 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6721 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6781 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6841 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6901 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6961 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7021 tttccccgaa aagtgccacc tgcagcctga atatgggcca aacaggatat ctgtggtaag
     7081 cagttcctgc cccggctcag ggccaagaac agatggaaca gctgaatatg ggccaaacag
     7141 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat
     7201 gcggtccagc cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga
     7261 cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc
     7321 gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc ggggcgccag
     7381 tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc
     7441 atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt
     7501 cagcgggggt ctttcatttg ggggctcgtc cgggatcggg agacccctgc ccagggacca
     7561 ccgacccacc accgggaggc aagctggcca gcaacttatc tgtgtctgtc cgattgtcta
     7621 gtgtctatga ctgattttat gcgcctgcgt cggtactagt tagctaacta gctctgtatc
     7681 tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc tgggagacgt
     7741 cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc gatgtggaat
     7801 ccgaccccgt caggatatgt ggttctggta ggagacgaga acctaaaaca gttcccgcct
     7861 ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct gctgcagcgc
     7921 tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc tgaaaattag
     7981 ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg
     8041 atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca
     8101 gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc
     8161 acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc
     8221 tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta
     8281 caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct
     8341 cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccggc
     8401 cggatcccag tgtggtggta ctaatagtaa tcaattacgg ggtcattagt tca
//
