LOCUS       sequence-272212-        2285 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 2285)
  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 2285)
  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-ePDZ-B1
FEATURES             Location/Qualifiers
     source          1..2285
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     532..554
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(592..610)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(743..1558)
                     /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(743..1558)
                     /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     1466..1485
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1550..1569
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1686..2273
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2175..2194
                     /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 tgggtcaccc gagttggggt tcagtatttc tggcggtgtt gggggtcgtg
      241 gtaatccctt ccgccctgac gatgacggaa tctttgtcac ccgtgtgcaa cccgaagggc
      301 ctgcaagtaa gttgttgcag ccaggcgata agatcattca ggctaatggc tattctttca
      361 tcaatattga gcatggtcaa gcggtatcct tattgaaaac ttttcaaaac acagtggaac
      421 ttatcattgt gcgtgaggtg ggtaacggcg ctaagcaaga gatccgtgtc cgtgtcgaaa
      481 aggatggcat tgcataaagc cccgacagta aaaagagacc ctgactagtc tccgggagct
      541 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gacacgaaag ggcctcgtga
      601 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggcc
      661 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
      721 gtatccgctc atgaattaat tcttagaaaa actcatcgag catcaaatga aactgcaatt
      781 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
      841 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
      901 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
      961 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt
     1021 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     1081 aaccgttatt cattcgtgat tgcgcctgag ccagacgaaa tacgcggtcg ctgttaaaag
     1141 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa caccgccagc gcatcaacaa
     1201 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc cctgggatcg
     1261 ccgtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     1321 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     1381 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga
     1441 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     1501 ccatgttgga atttaatcgc ggcctagagc aagacgtttc ccgttgaata tggctcataa
     1561 caccccttgt attactgttt atgtaagcag acagttttat tgtgaattct catgaccaaa
     1621 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1681 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1741 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1801 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     1861 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1921 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     1981 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2041 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2101 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2161 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2221 tgacttgagc gtcgattttt gtgatctcgt caggggggcg gagcctatgg aaaaacgcca
     2281 gcaac
//
