LOCUS       sequence-149738-        3542 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal tagging of human FANCF with 3xFLAG_Twin_Strep.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3542)
  AUTHORS   Dalvai M, Loehr J, Jacquet K, Huard CC, Roques C, Herst P, Cote J, 
            Doyon Y
  TITLE     A Scalable Genome-Editing-Based Approach for Mapping Multiprotein 
            Complexes in Human Cells.
  JOURNAL   Cell Rep. 2015 Oct 7. pii: S2211-1247(15)01020-7. doi: 
            10.1016/j.celrep.2015.09.009.
  PUBMED    26456817
REFERENCE   2  (bases 1 to 3542)
  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"; journalName: "Cell Rep. 
            2015 Oct 7. pii: S2211-1247(15)01020-7. doi: 
            10.1016/j.celrep.2015.09.009."
COMMENT     Sequence Label: FancF_N_FS_v2_Donor
FEATURES             Location/Qualifiers
     source          1..3542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     regulatory      565..574
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             586..651
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(661..666,667..690,691..726,727..750)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     primer_bind     complement(1552..1569)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1723..2311)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1803..1822)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2482..3273,3274..3342))
                     /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     3105..3124
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3343..3447)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3515..3533
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatggt accaacctca acacagattc tagttttagg caaagctcag aaaatttcta
      241 cttaaggata tttccaaagc gaaaggaagc gcggagacgt tcatgactgg catcatctcg
      301 cacgtggttc cggaaattct cggtaggatg ccctacatct gctctccctc cactaagaag
      361 aacctctttg tgtggcgaaa gtaaaagtat tagggctttt aagttgccca gagtcaagga
      421 acacggataa agacgctggg agattgacat gcatttcgac caatagcatt gcagagaggc
      481 gtatcatttc gcggatgttc caatcagtac gcagagagtc gccgtctcca aggtgaaagc
      541 ggaagtaggg ccttcgcgca cctcgccacc atgggatccg ccggcgacta caaggaccac
      601 gacggcgatt ataaggatca cgacatcgac tacaaagacg acgatgacaa gggcgccagc
      661 agcgcctggt cccaccctca gtttgagaag ggcggaggct ctggcggcgg aagcggagga
      721 tctgcttgga gccaccccca gttcgaaaag ggcgccgaat cccttctgca gcacctggat
      781 cgcttttccg agcttctggc ggtctcaagc actacctacg tcagcacctg ggaccccgcc
      841 accgtgcgcc gggccttgca gtgggcgcgc tacctgcgcc acatccatcg gcgctttggt
      901 cggcatggcc ccattcgcac ggctctggag cggcggctgc acaaccagtg gaggcaagag
      961 ggcggctttg ggcggggtcc agttccggga ttagcgaact tccaggccct cggtcactgt
     1021 gacgtcctgc tctctctgcg cctgctggag aaccgggccc tcggggatgc agctcgttac
     1081 cacctggtgc agcaactctt tcccggcccg ggcgtccggg acgccgatga ggagacactc
     1141 caagagagcc tggcccgcct tgcccgccgg cggtctgcgg tgcacatgct gcgcttcaat
     1201 ggctatagag agaacccaaa tctccaggag gactctctga tgaagaccca ggcggagctg
     1261 ctgctggagc gtctgcagga ggtggggaag gccgaagcgg agcgtcccgc caggtttctc
     1321 agcagcctgt gggagcgctt gcctcagaac aacttcctga aggtgatagc ggtggcgctg
     1381 ttgcagccgc ctttgtctcg tcggccccaa gaagagttgg aacccggcat ccacaaatca
     1441 cctggagagg ggagccaagt gctagtccac tggcttctgg ggaattctgc attaatgaat
     1501 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     1561 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     1621 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     1681 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1741 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     1801 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     1861 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     1921 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     1981 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2041 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2101 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2161 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2221 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2281 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2341 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2401 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2461 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     2521 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     2581 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     2641 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     2701 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     2761 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     2821 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     2881 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     2941 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3001 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3061 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     3121 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3181 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3241 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3301 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     3361 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3421 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     3481 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     3541 tc
//
