LOCUS       sequence-494872-        7541 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression construct for N-terminal S 
            protein-Flag-Streptavidin binding peptide (SFB)-tagged SLX4IP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7541)
  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 7541)
  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-SLX4IP
FEATURES             Location/Qualifiers
     source          1..7541
                     /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(2171..2195)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2378..2395)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     complement(2457..2477)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2462..2791
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2642..2777
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2801..3400
                     /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(2801..2820)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3297..3317
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3621..4090
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4220..4237)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4391..4979)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4471..4490)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5139..5930,5931..5999))
                     /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     5762..5781
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6000..6104)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             6135..6604
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    6669..6868
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             7069..7485
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     7415..7437
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     7453..7469
                     /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 tggcatctaa gaaatttgct
      961 gttaaatgtg ggaattttgc tgtcctcgtg gatcttcata tcttgccaca aggttcaaac
     1021 aaagatacaa gctggttttc tgaacagaag aaagaggaag tctgtttact gttaaaagaa
     1081 accattgatt caagagttca ggagtacttg gaagttcgca aacagcacag gccatcaaat
     1141 gcagaattca caagatccaa tcccttgtcc ttaaaaggtt atggctttca aatcacagcc
     1201 tatttcctca agagagggat acgccttcgc tgcatcagga gcacacagaa tgctgaactc
     1261 tgtgtattcc ctgacagatt tgtggtttgt gtcagtcagc ttgcattcag tcgtgatctt
     1321 ttagcaagtc agaatgaaga tttgacagaa agagttctcc atggagtgtc tgattacttt
     1381 gctgagtgtg cagagagttc acttcctccc agtgcaaagc tccggagaaa tgctctgaaa
     1441 gaaattgtga aaagaactga aacaaaaagc agtgtcacga gcaaatcgca gaccagaaga
     1501 gacactgtgg aaacatctag tgactcagtg attgcagaga tagcaaggag gaggaatgat
     1561 ggtcaggctt cctccagtcc cccatcagaa tccatgggac aagcaaagga ttccataaag
     1621 gcagctgaga gccactgggg gcttcctgtt caaaagctgg aaaaagttaa tcagacccag
     1681 ccagaagaca ctagtggcca gcaaaaacct catcctgggg agcggttaaa gacagggctt
     1741 ctaagcagga gccccgtctg tagctgtgag tcagcatcac catgtccaaa acaaagtcca
     1801 cgagtggcca aaacccaaca gaaacgcagg aactgcagct ctgcggaaga cttcgaccac
     1861 cacgggagag tttctcttgg aagtgatcga ttagtcccga gagaaataat agtggaaaaa
     1921 agcaaagctg tcagggtttt gccagcttca gagttgtcag atccagggtt acttttgaaa
     1981 caagatttgg caaaaaccac gtctaaggaa gagttgcatg ttttggaaag tctctcctcc
     2041 agacatctta tgaaaaataa cccagggcag gcacagcaaa ccggcttagc cacaaacact
     2101 gaaagattat ctacaattca gaacagccca accaagaaaa gaaagaaata cgaaagaggc
     2161 cattaataaa acccagcttt cttgtacaaa gtggctcgag cagggatccg cccctctccc
     2221 tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc
     2281 tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc
     2341 cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt
     2401 ctgttgaatg tcgtgaagga agcgtaggaa ttcgccagca cagtggtcga ccctgtggaa
     2461 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2521 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     2581 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     2641 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     2701 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     2761 aggctttttt ggaggcctag gcttttgcaa aaagcttacc atgaccgagt acaagcccac
     2821 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     2881 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     2941 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     3001 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     3061 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     3121 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     3181 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     3241 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     3301 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     3361 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga cgcccgcccc acgacccgca
     3421 gcgcccgacc gaaaggagcg cacgacccca tgcatcgata aaataaaaga ttttatttag
     3481 tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt
     3541 aacgccattt tgcaaggcat ggaaaaatac ataactgaga atagagaagt tcagatcaag
     3601 gtcaggaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc
     3661 ctgccccggc tcagggccaa gaacagatgg aacagctgaa tatgggccaa acaggatatc
     3721 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc
     3781 cagccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa
     3841 atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc
     3901 ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggggcg ccagtcctcc
     3961 gattgactga gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga
     4021 cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg
     4081 gggtctttca catgcagcat gtatcaaaat taatttggtt ttttttctta agtatttaca
     4141 ttaaatggcc atagttgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4201 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4261 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4321 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4381 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4441 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4501 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4561 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4621 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4681 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4741 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4801 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4861 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4921 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4981 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5041 ggattttggt catgagatta tcaaaaagga tcttcaccta gatccttttg cggccgcaaa
     5101 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5161 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5221 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5281 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5341 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5401 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5461 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5521 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5581 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5641 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5701 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     5761 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     5821 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     5881 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     5941 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6001 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6061 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6121 gtgccacctg cagcctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     6181 cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt
     6241 aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc ggtccagccc
     6301 tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc
     6361 ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc
     6421 tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga
     6481 ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg
     6541 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct
     6601 ttcatttggg ggctcgtccg ggatcgggag acccctgccc agggaccacc gacccaccac
     6661 cgggaggcaa gctggccagc aacttatctg tgtctgtccg attgtctagt gtctatgact
     6721 gattttatgc gcctgcgtcg gtactagtta gctaactagc tctgtatctg gcggacccgt
     6781 ggtggaactg acgagttctg aacacccggc cgcaaccctg ggagacgtcc cagggacttt
     6841 gggggccgtt tttgtggccc gacctgagga agggagtcga tgtggaatcc gaccccgtca
     6901 ggatatgtgg ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gtctgaattt
     6961 ttgctttcgg tttggaaccg aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt
     7021 ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg aaaattaggg ccagactgtt
     7081 accactccct taagtttgac cttaggtcac tggaaagatg tcgagcggat cgctcacaac
     7141 cagtcggtag atgtcaagaa gagacgttgg gttaccttct gctctgcaga atggccaacc
     7201 tttaacgtcg gatggccgcg agacggcacc tttaaccgag acctcatcac ccaggttaag
     7261 atcaaggtct tttcacctgg cccgcatgga cacccagacc aggtccccta catcgtgacc
     7321 tgggaagcct tggcttttga cccccctccc tgggtcaagc cctttgtaca ccctaagcct
     7381 ccgcctcctc ttcctccatc cgccccgtct ctcccccttg aacctcctcg ttcgaccccg
     7441 cctcgatcct ccctttatcc agccctcact ccttctctag gcgccggccg gatcccagtg
     7501 tggtggtact aatagtaatc aattacgggg tcattagttc a
//
