LOCUS       sequence-495254-        7541 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression construct for C-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: SLX4IP-SFB
FEATURES             Location/Qualifiers
     source          1..7541
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     464..484
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             788..1257
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(1387..1404)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1558..2146)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1638..1657)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2306..3097,3098..3166))
                     /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     2929..2948
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3167..3271)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             3302..3771
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    3836..4035
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             4236..4652
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     4582..4604
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     4620..4636
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        4730..5033
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5034..5237
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5188..5208
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             5297..5341
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             5375..5398
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5402..5425
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5435..5548
                     /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    5621..5645
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(6879..6903)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(7086..7103)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     complement(7165..7185)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7170..7499
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7350..7485
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(7509..7541,1..567)
                     /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(7509..7528)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
       61 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
      121 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
      181 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
      241 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
      301 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
      361 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
      421 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
      481 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
      541 tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa
      601 aggagcgcac gaccccatgc atcgataaaa taaaagattt tatttagtct ccagaaaaag
      661 gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc
      721 aaggcatgga aaaatacata actgagaata gagaagttca gatcaaggtc aggaacagat
      781 ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
      841 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
      901 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
      961 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
     1021 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
     1081 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
     1141 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
     1201 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcacat
     1261 gcagcatgta tcaaaattaa tttggttttt tttcttaagt atttacatta aatggccata
     1321 gttgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     1381 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     1441 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     1501 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1561 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1621 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1681 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1741 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1801 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     1861 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     1921 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     1981 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2041 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2101 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2161 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2221 gagattatca aaaaggatct tcacctagat ccttttgcgg ccgcaaatca atctaaagta
     2281 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     2341 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     2401 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     2461 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     2521 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     2581 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     2641 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     2701 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     2761 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     2821 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     2881 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     2941 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     3001 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     3061 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     3121 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     3181 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     3241 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgcag
     3301 cctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     3361 agaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc
     3421 cccggctcag ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta
     3481 gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt
     3541 gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa
     3601 taaaagagcc cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg
     3661 gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc
     3721 ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc atttgggggc
     3781 tcgtccggga tcgggagacc cctgcccagg gaccaccgac ccaccaccgg gaggcaagct
     3841 ggccagcaac ttatctgtgt ctgtccgatt gtctagtgtc tatgactgat tttatgcgcc
     3901 tgcgtcggta ctagttagct aactagctct gtatctggcg gacccgtggt ggaactgacg
     3961 agttctgaac acccggccgc aaccctggga gacgtcccag ggactttggg ggccgttttt
     4021 gtggcccgac ctgaggaagg gagtcgatgt ggaatccgac cccgtcagga tatgtggttc
     4081 tggtaggaga cgagaaccta aaacagttcc cgcctccgtc tgaatttttg ctttcggttt
     4141 ggaaccgaag ccgcgcgtct tgtctgctgc agcgctgcag catcgttctg tgttgtctct
     4201 gtctgactgt gtttctgtat ttgtctgaaa attagggcca gactgttacc actcccttaa
     4261 gtttgacctt aggtcactgg aaagatgtcg agcggatcgc tcacaaccag tcggtagatg
     4321 tcaagaagag acgttgggtt accttctgct ctgcagaatg gccaaccttt aacgtcggat
     4381 ggccgcgaga cggcaccttt aaccgagacc tcatcaccca ggttaagatc aaggtctttt
     4441 cacctggccc gcatggacac ccagaccagg tcccctacat cgtgacctgg gaagccttgg
     4501 cttttgaccc ccctccctgg gtcaagccct ttgtacaccc taagcctccg cctcctcttc
     4561 ctccatccgc cccgtctctc ccccttgaac ctcctcgttc gaccccgcct cgatcctccc
     4621 tttatccagc cctcactcct tctctaggcg ccggccggat cccagtgtgg tggtactaat
     4681 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     4741 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     4801 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     4861 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     4921 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     4981 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     5041 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     5101 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     5161 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     5221 tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac cggactcaga
     5281 tctcgaggcc accatgaaag aaaccgctgc tgctaaattc gaacgccagc acatggacag
     5341 cggagccggc gcaggagccg gcgctgacgc gcctgactac aaagacgatg acgacaaggg
     5401 agattacaag gatgacgatg acaaaggcgc gtcgatggac gagaagacca ccggctggcg
     5461 gggcggccac gtggtggagg gcctggccgg cgagctggag cagctgcggg ccaggctgga
     5521 gcaccaccct cagggccagc gggagccctc cggcggctgc aagctgggcg gcgcgcctga
     5581 attctgcagt cgacggtacc gcgggcccat cgctcgagag acaagtttgt acaaaaaagc
     5641 aggcttcatg gcatctaaga aatttgctgt taaatgtggg aattttgctg tcctcgtgga
     5701 tcttcatatc ttgccacaag gttcaaacaa agatacaagc tggttttctg aacagaagaa
     5761 agaggaagtc tgtttactgt taaaagaaac cattgattca agagttcagg agtacttgga
     5821 agttcgcaaa cagcacaggc catcaaatgc agaattcaca agatccaatc ccttgtcctt
     5881 aaaaggttat ggctttcaaa tcacagccta tttcctcaag agagggatac gccttcgctg
     5941 catcaggagc acacagaatg ctgaactctg tgtattccct gacagatttg tggtttgtgt
     6001 cagtcagctt gcattcagtc gtgatctttt agcaagtcag aatgaagatt tgacagaaag
     6061 agttctccat ggagtgtctg attactttgc tgagtgtgca gagagttcac ttcctcccag
     6121 tgcaaagctc cggagaaatg ctctgaaaga aattgtgaaa agaactgaaa caaaaagcag
     6181 tgtcacgagc aaatcgcaga ccagaagaga cactgtggaa acatctagtg actcagtgat
     6241 tgcagagata gcaaggagga ggaatgatgg tcaggcttcc tccagtcccc catcagaatc
     6301 catgggacaa gcaaaggatt ccataaaggc agctgagagc cactgggggc ttcctgttca
     6361 aaagctggaa aaagttaatc agacccagcc agaagacact agtggccagc aaaaacctca
     6421 tcctggggag cggttaaaga cagggcttct aagcaggagc cccgtctgta gctgtgagtc
     6481 agcatcacca tgtccaaaac aaagtccacg agtggccaaa acccaacaga aacgcaggaa
     6541 ctgcagctct gcggaagact tcgaccacca cgggagagtt tctcttggaa gtgatcgatt
     6601 agtcccgaga gaaataatag tggaaaaaag caaagctgtc agggttttgc cagcttcaga
     6661 gttgtcagat ccagggttac ttttgaaaca agatttggca aaaaccacgt ctaaggaaga
     6721 gttgcatgtt ttggaaagtc tctcctccag acatcttatg aaaaataacc cagggcaggc
     6781 acagcaaacc ggcttagcca caaacactga aagattatct acaattcaga acagcccaac
     6841 caagaaaaga aagaaatacg aaagaggcca ttaataaaac ccagctttct tgtacaaagt
     6901 ggctcgagca gggatccgcc cctctccctc ccccccccct aacgttactg gccgaagccg
     6961 cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt
     7021 tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct
     7081 ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cgtaggaatt
     7141 cgccagcaca gtggtcgacc ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     7201 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     7261 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     7321 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     7381 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     7441 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     7501 agcttaccat gaccgagtac aagcccacgg tgcgcctcgc c
//
