LOCUS       sequence-494917-        8753 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression construct for N-terminal S 
            protein-Flag-Streptavidin binding peptide (SFB)-tagged XPF D814-916.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8753)
  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 8753)
  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-D814-916
FEATURES             Location/Qualifiers
     source          1..8753
                     /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(3383..3407)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3590..3607)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     complement(3669..3689)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3674..4003
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3854..3989
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4013..4612
                     /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(4013..4032)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4509..4529
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4833..5302
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5432..5449)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5603..6191)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5683..5702)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6351..7142,7143..7211))
                     /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     6974..6993
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7212..7316)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             7347..7816
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    7881..8080
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             8281..8697
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     8627..8649
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     8665..8681
                     /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 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 agttgttttg acacccagct ttcttgtaca aagtggctcg agcagggatc
     3421 cgcccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg
     3481 tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc
     3541 cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa
     3601 ggaatgcaag gtctgttgaa tgtcgtgaag gaagcgtagg aattcgccag cacagtggtc
     3661 gaccctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3721 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3781 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3841 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3901 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     3961 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctta ccatgaccga
     4021 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
     4081 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
     4141 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     4201 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     4261 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     4321 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     4381 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     4441 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     4501 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     4561 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacgcccgcc
     4621 ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catgcatcga taaaataaaa
     4681 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa
     4741 gctagcttaa gtaacgccat tttgcaaggc atggaaaaat acataactga gaatagagaa
     4801 gttcagatca aggtcaggaa cagatggaac agctgaatat gggccaaaca ggatatctgt
     4861 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggaacagctg aatatgggcc
     4921 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc
     4981 ccagatgcgg tccagccctc agcagtttct agagaaccat cagatgtttc cagggtgccc
     5041 caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt
     5101 ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcgggg
     5161 cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc
     5221 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
     5281 acccgtcagc gggggtcttt cacatgcagc atgtatcaaa attaatttgg ttttttttct
     5341 taagtattta cattaaatgg ccatagttgc attaatgaat cggccaacgc gcggggagag
     5401 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5461 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5521 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5581 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5641 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5701 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5761 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5821 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5881 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5941 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6001 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6061 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6121 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6181 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6241 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6301 tgcggccgca aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6361 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6421 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6481 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6541 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6601 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6661 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6721 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6781 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6841 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6901 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6961 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7021 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7081 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7141 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7201 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7261 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7321 tttccccgaa aagtgccacc tgcagcctga atatgggcca aacaggatat ctgtggtaag
     7381 cagttcctgc cccggctcag ggccaagaac agatggaaca gctgaatatg ggccaaacag
     7441 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat
     7501 gcggtccagc cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga
     7561 cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc
     7621 gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc ggggcgccag
     7681 tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc
     7741 atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt
     7801 cagcgggggt ctttcatttg ggggctcgtc cgggatcggg agacccctgc ccagggacca
     7861 ccgacccacc accgggaggc aagctggcca gcaacttatc tgtgtctgtc cgattgtcta
     7921 gtgtctatga ctgattttat gcgcctgcgt cggtactagt tagctaacta gctctgtatc
     7981 tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc tgggagacgt
     8041 cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc gatgtggaat
     8101 ccgaccccgt caggatatgt ggttctggta ggagacgaga acctaaaaca gttcccgcct
     8161 ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct gctgcagcgc
     8221 tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc tgaaaattag
     8281 ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg
     8341 atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca
     8401 gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc
     8461 acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc
     8521 tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta
     8581 caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct
     8641 cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccggc
     8701 cggatcccag tgtggtggta ctaatagtaa tcaattacgg ggtcattagt tca
//
