LOCUS       sequence-494934-        8303 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression construct for N-terminal S 
            protein-Flag-Streptavidin binding peptide (SFB)-tagged XPF D205-457.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8303)
  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 8303)
  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-D205-457
FEATURES             Location/Qualifiers
     source          1..8303
                     /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(2933..2957)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3140..3157)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     complement(3219..3239)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3224..3553
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3404..3539
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3563..4162
                     /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(3563..3582)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4059..4079
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4383..4852
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4982..4999)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5153..5741)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5233..5252)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5901..6692,6693..6761))
                     /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     6524..6543
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6762..6866)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             6897..7366
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    7431..7630
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             7831..8247
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     8177..8199
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     8215..8231
                     /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 aagttcaaag
     1561 agaattagga aatctcacaa aagacctaaa gacccccaaa acaaagaacg ggcttctacc
     1621 aaagaaagaa ccctcaaaaa gaaaaaacgg aagttgacct taactcaaat ggtaggaaaa
     1681 cctgaagaac tggaagagga aggagatgtc gaggaaggat atcgtcgaga aataagcagt
     1741 agcccagaaa gctgcccgga agaaattaag catgaagaat ttgatgtaaa tttgtcatcg
     1801 gatgctgctt tcggaatcct gaaagaaccc ctcactatca tccatccgct tctgggttgc
     1861 agcgacccct atgctctgac aagggtacta catgaagtgg agccaagata cgtggttctt
     1921 tatgacgcag agctaacctt tgttcggcag cttgaaattt acagggcgag taggcctggg
     1981 aaacctctga gggtttactt tcttatatac ggaggttcaa ctgaggaaca acgctatctc
     2041 actgctttgc ggaaagaaaa ggaagctttt gaaaaactca taagggaaaa agcaagcatg
     2101 gttgtccctg aagaaagaga aggcagagat gaaacaaact tagacctagt aagaggcaca
     2161 gcatctgcag atgtttccac tgacactcgg aaagccggtg gccaggaaca gaatggtaca
     2221 cagcaaagca tagttgtgga tatgcgtgaa tttcgaagtg agcttccatc tctgatccat
     2281 cgtcggggca ttgacattga acccgtgact ttagaggttg gagattacat cctcactcca
     2341 gaaatgtgcg tggagcgcaa gagtatcagt gatttaatcg gctctttaaa taacggccgc
     2401 ctctacagcc agtgcatctc catgtcccgc tactacaagc gtcccgtgct tctgattgag
     2461 tttgacccta gcaagccttt ctctctcact tcccgaggtg ccttgtttca ggagatctcc
     2521 agcaatgaca ttagttccaa actcactctt cttacacttc acttccccag actacggatt
     2581 ctctggtgcc cctctcctca tgcaacggcg gagttgtttg aggagctgaa acaaagcaag
     2641 ccacagcctg atgcggcgac agcactggcc attacagcag attctgaaac ccttcccgag
     2701 tcagagaagt ataatcctgg tccccaagac ttcttgttaa aaatgccagg ggtgaatgcc
     2761 aaaaactgcc gctccttgat gcaccacgtt aagaacatcg cagaattagc agccctgtca
     2821 caagacgagc tcacgagtat tctggggaat gctgcaaatg ccaaacagct ttatgatttc
     2881 attcacacct cttttgcaga agtcgtatca aaaggaaaag ggaaaaagtg aaacccagct
     2941 ttcttgtaca aagtggctcg agcagggatc cgcccctctc cctccccccc ccctaacgtt
     3001 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc
     3061 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc
     3121 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag
     3181 gaagcgtagg aattcgccag cacagtggtc gaccctgtgg aatgtgtgtc agttagggtg
     3241 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3301 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3361 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3421 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3481 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3541 aggcttttgc aaaaagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
     3601 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     3661 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     3721 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     3781 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     3841 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     3901 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     3961 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     4021 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     4081 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     4141 gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag
     4201 cgcacgaccc catgcatcga taaaataaaa gattttattt agtctccaga aaaagggggg
     4261 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
     4321 atggaaaaat acataactga gaatagagaa gttcagatca aggtcaggaa cagatggaac
     4381 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     4441 aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg
     4501 ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct
     4561 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
     4621 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
     4681 ataaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact gagtcgcccg
     4741 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc
     4801 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt cacatgcagc
     4861 atgtatcaaa attaatttgg ttttttttct taagtattta cattaaatgg ccatagttgc
     4921 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4981 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5041 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5101 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5161 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5221 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5281 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5341 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5401 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5461 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5521 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5581 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5641 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5701 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5761 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5821 tatcaaaaag gatcttcacc tagatccttt tgcggccgca aatcaatcta aagtatatat
     5881 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5941 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6001 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6061 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6121 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6181 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6241 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6301 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6361 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6421 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6481 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6541 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6601 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6661 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6721 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6781 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6841 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgcagcctga
     6901 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac
     6961 agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
     7021 ctcagggcca agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagagaa
     7081 ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact
     7141 aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa
     7201 gagcccacaa cccctcactc ggggcgccag tcctccgatt gactgagtcg cccgggtacc
     7261 cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg
     7321 agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcatttg ggggctcgtc
     7381 cgggatcggg agacccctgc ccagggacca ccgacccacc accgggaggc aagctggcca
     7441 gcaacttatc tgtgtctgtc cgattgtcta gtgtctatga ctgattttat gcgcctgcgt
     7501 cggtactagt tagctaacta gctctgtatc tggcggaccc gtggtggaac tgacgagttc
     7561 tgaacacccg gccgcaaccc tgggagacgt cccagggact ttgggggccg tttttgtggc
     7621 ccgacctgag gaagggagtc gatgtggaat ccgaccccgt caggatatgt ggttctggta
     7681 ggagacgaga acctaaaaca gttcccgcct ccgtctgaat ttttgctttc ggtttggaac
     7741 cgaagccgcg cgtcttgtct gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg
     7801 actgtgtttc tgtatttgtc tgaaaattag ggccagactg ttaccactcc cttaagtttg
     7861 accttaggtc actggaaaga tgtcgagcgg atcgctcaca accagtcggt agatgtcaag
     7921 aagagacgtt gggttacctt ctgctctgca gaatggccaa cctttaacgt cggatggccg
     7981 cgagacggca cctttaaccg agacctcatc acccaggtta agatcaaggt cttttcacct
     8041 ggcccgcatg gacacccaga ccaggtcccc tacatcgtga cctgggaagc cttggctttt
     8101 gacccccctc cctgggtcaa gccctttgta caccctaagc ctccgcctcc tcttcctcca
     8161 tccgccccgt ctctccccct tgaacctcct cgttcgaccc cgcctcgatc ctccctttat
     8221 ccagccctca ctccttctct aggcgccggc cggatcccag tgtggtggta ctaatagtaa
     8281 tcaattacgg ggtcattagt tca
//
