LOCUS       sequence-272117-        1996 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 1996)
  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 1996)
  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(TP)4TG
FEATURES             Location/Qualifiers
     source          1..1996
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     243..265
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(303..321)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(454..1269)
                     /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(454..1269)
                     /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     1177..1196
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1261..1280
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1397..1984
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1886..1905
                     /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 gcagtgggta cccctacgcc
      181 aactcctaca cccactgggc attgcatcca gccccgagca aaaggagacc cctgactagt
      241 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgacacgaaa
      301 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
      361 gtcaggtggc cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
      421 cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga gcatcaaatg
      481 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
      541 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
      601 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
      661 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt
      721 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
      781 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gccagacgaa atacgcggtc
      841 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acaccgccag
      901 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
      961 ccctgggatc gccgtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     1021 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     1081 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     1141 caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     1201 taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt cccgttgaat
     1261 atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta ttgtgaattc
     1321 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1381 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1441 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1501 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     1561 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1621 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1681 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1741 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     1801 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     1861 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     1921 ttcgccacct ctgacttgag cgtcgatttt tgtgatctcg tcaggggggc ggagcctatg
     1981 gaaaaacgcc agcaac
//
