LOCUS       sequence-272196-        1966 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 1966)
  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 1966)
  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: pL2-G
FEATURES             Location/Qualifiers
     source          1..1966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     213..235
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(273..291)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(424..1239)
                     /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(424..1239)
                     /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     1147..1166
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1231..1250
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1367..1954
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1856..1875
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
       61 ttatcccctg attctgtgga taaccgtatt accgccaatt ggctcgccgc agccgaacga
      121 ccgagcgcag cgagtcaccc gaagaccatt ttggacgcca gcagtggggc attgcatcca
      181 gccccgagca aaaggagacc cctgactagt ctccgggagc tgcatgtgtc agaggttttc
      241 accgtcatca ccgaaacgcg cgacacgaaa gggcctcgtg atacgcctat ttttataggt
      301 taatgtcatg ataataatgg tttcttagac gtcaggtggc cttttcgggg aaatgtgcgc
      361 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgaattaa
      421 ttcttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
      481 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
      541 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
      601 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
      661 gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg
      721 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
      781 ttgcgcctga gccagacgaa atacgcggtc gctgttaaaa ggacaattac aaacaggaat
      841 cgaatgcaac cggcgcagga acaccgccag cgcatcaaca atattttcac ctgaatcagg
      901 atattcttct aatacctgga atgctgtttt ccctgggatc gccgtggtga gtaaccatgc
      961 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
     1021 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     1081 aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc
     1141 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
     1201 cggcctagag caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt
     1261 tatgtaagca gacagtttta ttgtgaattc tcatgaccaa aatcccttaa cgtgagtttt
     1321 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     1381 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     1441 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     1501 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     1561 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     1621 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     1681 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     1741 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     1801 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     1861 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     1921 tgtgatctcg tcaggggggc ggagcctatg gaaaaacgcc agcaac
//
