LOCUS       sequence-272111-        2008 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 2008)
  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 2008)
  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-(GGS)4GSG
FEATURES             Location/Qualifiers
     source          1..2008
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     255..277
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(315..333)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(466..1281)
                     /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(466..1281)
                     /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     1189..1208
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1273..1292
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1409..1996
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1898..1917
                     /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 gcagtgggtg gatccggagg
      181 tagcggtgga agtggaggat ctggatcagg gcattgcatc cagccccgag caaaaggaga
      241 cccctgacta gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
      301 cgcgacacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
      361 ggtttcttag acgtcaggtg gccttttcgg ggaaatgtgc gcggaacccc tatttgttta
      421 tttttctaaa tacattcaaa tatgtatccg ctcatgaatt aattcttaga aaaactcatc
      481 gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa
      541 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
      601 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
      661 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
      721 tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
      781 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagccagacg
      841 aaatacgcgg tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
      901 gaacaccgcc agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg
      961 gaatgctgtt ttccctggga tcgccgtggt gagtaaccat gcatcatcag gagtacggat
     1021 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc
     1081 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc
     1141 gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
     1201 tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctag agcaagacgt
     1261 ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag cagacagttt
     1321 tattgtgaat tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     1381 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     1441 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     1501 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     1561 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     1621 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     1681 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     1741 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     1801 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     1861 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     1921 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatct cgtcaggggg
     1981 gcggagccta tggaaaaacg ccagcaac
//
