LOCUS       sequence-272108-        1975 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 1975)
  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 1975)
  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-GGSG
FEATURES             Location/Qualifiers
     source          1..1975
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     222..244
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(282..300)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(433..1248)
                     /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(433..1248)
                     /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     1156..1175
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1240..1259
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1376..1963
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1865..1884
                     /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 gaagcgggca
      181 ttgcatccag ccccgagcaa aaggagaccc ctgactagtc tccgggagct gcatgtgtca
      241 gaggttttca ccgtcatcac cgaaacgcgc gacacgaaag ggcctcgtga tacgcctatt
      301 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggcc ttttcgggga
      361 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
      421 atgaattaat tcttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc
      481 aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc
      541 gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac
      601 atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc
      661 atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg
      721 ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt
      781 cattcgtgat tgcgcctgag ccagacgaaa tacgcggtcg ctgttaaaag gacaattaca
      841 aacaggaatc gaatgcaacc ggcgcaggaa caccgccagc gcatcaacaa tattttcacc
      901 tgaatcagga tattcttcta atacctggaa tgctgttttc cctgggatcg ccgtggtgag
      961 taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc
     1021 cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc
     1081 atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc
     1141 tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga
     1201 atttaatcgc ggcctagagc aagacgtttc ccgttgaata tggctcataa caccccttgt
     1261 attactgttt atgtaagcag acagttttat tgtgaattct catgaccaaa atcccttaac
     1321 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     1381 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     1441 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     1501 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     1561 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     1621 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     1681 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     1741 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     1801 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     1861 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     1921 gtcgattttt gtgatctcgt caggggggcg gagcctatgg aaaaacgcca gcaac
//
