LOCUS       sequence-272201-        2267 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 2267)
  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 2267)
  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-FRB
FEATURES             Location/Qualifiers
     source          1..2267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     514..536
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(574..592)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(725..1540)
                     /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(725..1540)
                     /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     1448..1467
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1532..1551
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1668..2255
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2157..2176
                     /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 tgggattctg tggcacgaga tgtggcatga gggcctggaa gaggcatctc
      241 gtctgtattt tggtgaacgc aacgttaaag gtatgttcga ggttttggag ccacttcacg
      301 ctatgatgga gcgcggtcca cagaccttga aagaaacctc gtttaatcaa gcgtatggcc
      361 gtgatctgat ggaggcacag gagtggtgtc gtaagtacat gaagagtggg aacgtaaagg
      421 acctgactca ggcgtgggat ctgtattacc acgtgttccg tcgcattggc attgcataaa
      481 gccccgacag taaaaagaga ccctgactag tctccgggag ctgcatgtgt cagaggtttt
      541 caccgtcatc accgaaacgc gcgacacgaa agggcctcgt gatacgccta tttttatagg
      601 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg ccttttcggg gaaatgtgcg
      661 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta
      721 attcttagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
      781 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
      841 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
      901 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
      961 cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag
     1021 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     1081 attgcgcctg agccagacga aatacgcggt cgctgttaaa aggacaatta caaacaggaa
     1141 tcgaatgcaa ccggcgcagg aacaccgcca gcgcatcaac aatattttca cctgaatcag
     1201 gatattcttc taatacctgg aatgctgttt tccctgggat cgccgtggtg agtaaccatg
     1261 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     1321 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     1381 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     1441 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     1501 gcggcctaga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     1561 ttatgtaagc agacagtttt attgtgaatt ctcatgacca aaatccctta acgtgagttt
     1621 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     1681 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     1741 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     1801 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     1861 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     1921 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     1981 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     2041 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     2101 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     2161 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     2221 ttgtgatctc gtcagggggg cggagcctat ggaaaaacgc cagcaac
//
