LOCUS       sequence-272099-        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-GSPAG
FEATURES             Location/Qualifiers
     source          1..1978
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     225..247
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(285..303)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(436..1251)
                     /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(436..1251)
                     /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     1159..1178
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1243..1262
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1379..1966
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1868..1887
                     /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 gcagtgggtt cacctgcagg
      181 gcattgcatc cagccccgag caaaaggaga cccctgacta gtctccggga gctgcatgtg
      241 tcagaggttt tcaccgtcat caccgaaacg cgcgacacga aagggcctcg tgatacgcct
      301 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gccttttcgg
      361 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      421 ctcatgaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
      481 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
      541 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
      601 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
      661 accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac
      721 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
      781 attcattcgt gattgcgcct gagccagacg aaatacgcgg tcgctgttaa aaggacaatt
      841 acaaacagga atcgaatgca accggcgcag gaacaccgcc agcgcatcaa caatattttc
      901 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttccctggga tcgccgtggt
      961 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     1021 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     1081 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     1141 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     1201 ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca taacacccct
     1261 tgtattactg tttatgtaag cagacagttt tattgtgaat tctcatgacc aaaatccctt
     1321 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     1381 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     1441 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     1501 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     1561 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     1621 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     1681 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     1741 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     1801 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     1861 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     1921 agcgtcgatt tttgtgatct cgtcaggggg gcggagccta tggaaaaacg ccagcaac
//
