LOCUS       sequence-272105-        1978 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 1978)
  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 1978)
  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-GPPPG
FEATURES             Location/Qualifiers
     source          1..1978
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      131..718
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     620..639
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     955..977
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1015..1033)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(join(1166..1978,1..3))
                     /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(join(1166..1978,1..3))
                     /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     1889..1908
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     join(1973..1978,1..14)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 cataacaccc cttgtattac tgtttatgta agcagacagt tttattgtga attctcatga
       61 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
      121 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
      181 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
      241 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
      301 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
      361 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
      421 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
      481 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
      541 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
      601 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
      661 acctctgact tgagcgtcga tttttgtgat ctcgtcaggg gggcggagcc tatggaaaaa
      721 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
      781 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccaatt ggctcgccgc
      841 agccgaacga ccgagcgcag cgagtcaccc gaagaccatt ttggacgcca gcagtgggtc
      901 cgcctccagg gcattgcatc cagccccgag caaaaggaga cccctgacta gtctccggga
      961 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgacacga aagggcctcg
     1021 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     1081 gccttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     1141 tatgtatccg ctcatgaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca
     1201 atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag
     1261 gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc
     1321 cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa
     1381 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt
     1441 ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa
     1501 ccaaaccgtt attcattcgt gattgcgcct gagccagacg aaatacgcgg tcgctgttaa
     1561 aaggacaatt acaaacagga atcgaatgca accggcgcag gaacaccgcc agcgcatcaa
     1621 caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt ttccctggga
     1681 tcgccgtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa
     1741 gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa
     1801 cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat
     1861 agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag
     1921 catccatgtt ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggct
//
