LOCUS       sequence-272202-        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-G(P)10G
FEATURES             Location/Qualifiers
     source          1..1999
                     /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     976..998
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1036..1054)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(join(1187..1999,1..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(1187..1999,1..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     1910..1929
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     join(1994..1999,1..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 cgcctccacc acctccgcca cctcctccag ggcattgcat ccagccccga gcaaaaggag
      961 acccctgact agtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     1021 gcgcgacacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     1081 tggtttctta gacgtcaggt ggccttttcg gggaaatgtg cgcggaaccc ctatttgttt
     1141 atttttctaa atacattcaa atatgtatcc gctcatgaat taattcttag aaaaactcat
     1201 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     1261 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     1321 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     1381 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     1441 atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     1501 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagccagac
     1561 gaaatacgcg gtcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
     1621 ggaacaccgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
     1681 ggaatgctgt tttccctggg atcgccgtgg tgagtaacca tgcatcatca ggagtacgga
     1741 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
     1801 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
     1861 cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc
     1921 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggccta gagcaagacg
     1981 tttcccgttg aatatggct
//
