LOCUS       sequence-272204-        1999 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 1999)
  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 1999)
  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-GGASPAAPAPAG
FEATURES             Location/Qualifiers
     source          1..1999
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     246..268
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(306..324)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(457..1272)
                     /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(457..1272)
                     /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     1180..1199
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1264..1283
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1400..1987
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1889..1908
                     /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 gagcatctcc
      181 tgcagctcca gctcctgcag ggcattgcat ccagccccga gcaaaaggag acccctgact
      241 agtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgacacg
      301 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
      361 gacgtcaggt ggccttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
      421 atacattcaa atatgtatcc gctcatgaat taattcttag aaaaactcat cgagcatcaa
      481 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
      541 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
      601 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
      661 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
      721 tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
      781 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagccagac gaaatacgcg
      841 gtcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacaccgc
      901 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
      961 tttccctggg atcgccgtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     1021 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     1081 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     1141 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     1201 atataaatca gcatccatgt tggaatttaa tcgcggccta gagcaagacg tttcccgttg
     1261 aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgtgaa
     1321 ttctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1381 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1441 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1501 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     1561 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1621 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1681 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1741 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1801 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1861 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1921 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatc tcgtcagggg ggcggagcct
     1981 atggaaaaac gccagcaac
//
