LOCUS       sequence-272191-        3188 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  iFLinkC Linker cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3188)
  AUTHORS   Grawe A, Ranglack J, Weyrich A, Stein V
  TITLE     iFLinkC: an iterative functional linker cloning strategy for the 
            combinatorial assembly and recombination of linker peptides with 
            functional domains.
  JOURNAL   Nucleic Acids Res. 2020 Jan 11. pii: 5700546. doi: 
            10.1093/nar/gkz1210.
  PUBMED    31925441
REFERENCE   2  (bases 1 to 3188)
  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: "Nucleic 
            Acids Res. 2020 Jan 11. pii: 5700546. doi: 10.1093/nar/gkz1210."
COMMENT     Sequence Label: pFD-MBP-CS(TEV)-StrepTag-II-AI(TVMV)
FEATURES             Location/Qualifiers
     source          1..3188
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             195..1292
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1266..1289
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1305..1325
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1332..1355
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     1435..1457
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1495..1513)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(1646..2461)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1646..2461)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2369..2388
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2453..2472
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2589..3176
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3078..3097
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
       61 ttatcccctg attctgtgga taaccgtatt accgccaatt gagtgagctg ataccgctcg
      121 ccgcagccga acgaccgagc gcagcgagtc acccgcgaaa tacaagaaga caattttccc
      181 gcaccagcag tgggaaaatc gaagaaggta aactggtaat ctggattaac ggcgataaag
      241 gctataacgg tctggctgaa gtcggtaaga aattcgagaa agataccgga attaaagtca
      301 ccgttgagca tccggataaa ctggaagaga aattcccaca ggttgcggca actggcgatg
      361 gccctgacat tatcttctgg gcacacgacc gctttggtgg ctacgctcaa tctggcctgt
      421 tggctgaaat caccccggac aaagcgttcc aggacaagct gtatccgttt acctgggatg
      481 ccgtacgtta caacggcaag ctgattgctt acccgatcgc tgttgaagcg ttatcgctga
      541 tttataacaa agatctgctg ccgaacccgc caaaaacctg ggaagagatc ccggcgctgg
      601 ataaagaact gaaagcgaaa ggtaagagcg cgctgatgtt caacctgcaa gaaccgtact
      661 tcacctggcc gctgattgct gctgacgggg gttatgcgtt caagtatgaa aacggcaagt
      721 acgacattaa agacgtgggc gtggataacg ctggcgcgaa agcgggtctg accttcctgg
      781 ttgacctgat taaaaacaaa cacatgaatg cagacaccga ttactccatc gcagaagctg
      841 cctttaataa aggcgaaaca gcgatgacca tcaacggccc gtgggcatgg tccaacatcg
      901 acaccagcaa agtgaattat ggtgtaacgg tactgccgac cttcaagggt caaccatcca
      961 aaccgttcgt tggcgtgctg agcgcaggta ttaacgccgc cagtccgaac aaagagctgg
     1021 caaaagagtt cctcgaaaac tatctgctga ctgatgaagg tctggaagcg gttaataaag
     1081 acaaaccgct gggtgccgta gcgctgaagt cttacgagga agagttggtg aaagatccgc
     1141 gtattgccgc cactatggaa aacgcccaga aaggtgaaat catgccgaac atcccgcaga
     1201 tgtccgcttt ctggtatgcc gtgcgtactg cggtgatcaa cgccgccagc ggtcgtcaga
     1261 ctgtcgatga agccctgaaa gacgcgcaga ctaattcgag ctcggagaac ctgtatttcc
     1321 agtccagtgg ctggagccac ccgcagttcg aaaagtcagg aggtcgcgaa tatgtgcgct
     1381 ttgcgccggg cattgcataa agccccgaca gtaaaaagag accctgacta gtctccggga
     1441 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgacacga aagggcctcg
     1501 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     1561 gccttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     1621 tatgtatccg ctcatgaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca
     1681 atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag
     1741 gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc
     1801 cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa
     1861 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt
     1921 ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa
     1981 ccaaaccgtt attcattcgt gattgcgcct gagccagacg aaatacgcgg tcgctgttaa
     2041 aaggacaatt acaaacagga atcgaatgca accggcgcag gaacaccgcc agcgcatcaa
     2101 caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt ttccctggga
     2161 tcgccgtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa
     2221 gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa
     2281 cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat
     2341 agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag
     2401 catccatgtt ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca
     2461 taacacccct tgtattactg tttatgtaag cagacagttt tattgtgaat tctcatgacc
     2521 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     2581 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     2641 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     2701 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     2761 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     2821 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     2881 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     2941 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     3001 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3061 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3121 ctctgacttg agcgtcgatt tttgtgatct cgtcaggggg gcggagccta tggaaaaacg
     3181 ccagcaac
//
