LOCUS       sequence-272189-        2660 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 2660)
  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 2660)
  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: pFD-NIa-Protease-TVMV
FEATURES             Location/Qualifiers
     source          1..2660
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     907..929
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(967..985)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(1118..1933)
                     /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(1118..1933)
                     /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     1841..1860
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1925..1944
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2061..2648
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2550..2569
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
       61 ttatcccctg attctgtgga taaccgtatt accgccaatt gagtgagctg ataccgctcg
      121 ccgcagccga acgaccgagc gcagcgagtc acccgcgaaa tacaagaaga caattttccc
      181 gcaccagcag tgggaccaag gcgcttctga aaggcgtgcg tgacttcaat cctatttctg
      241 cctgtgtatg tttgttagag aacagttctg acggccactc tgaaaggcta ttcggtatag
      301 gcttcggacc gtacatcatc gccaaccaac acctatttag aagaaataac ggcgaactga
      361 ccatcaaaac catgcacggt gagttcaaag ttaagaactc cactcagctt cagatgaaac
      421 ctgttgaagg aagagatatt atagtgatta agatggcaaa agactttccg ccatttccac
      481 aaaaattgaa atttcgtcag ccgaccataa aagatagagt ttgtatggta agtactaact
      541 ttcaacagaa atcagtctca agtcttgtta gcgaaagtag tcacattgtg cacaaagagg
      601 atactagctt ttggcagcac tggataacta cgaaagacgg tcagtgtggt agtccattgg
      661 tatcaattat agacggaaat atattaggta tccacagtct aactcatact accaatggtt
      721 ccaattattt cgtcgagttt cctgaaaagt ttgtggcgac ttatctggac gcagccgacg
      781 gatggtgcaa aaactggaag tttaacgcag acaaaataag ttggggatca ttcaccttag
      841 ttgaggatgc accggaggac ggcattgcat aaagccccga cagtaaaaag agaccctgac
      901 tagtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgacac
      961 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     1021 agacgtcagg tggccttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     1081 aatacattca aatatgtatc cgctcatgaa ttaattctta gaaaaactca tcgagcatca
     1141 aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt
     1201 tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc
     1261 ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa
     1321 taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa
     1381 gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat
     1441 cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagccaga cgaaatacgc
     1501 ggtcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacaccg
     1561 ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg
     1621 ttttccctgg gatcgccgtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct
     1681 tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa
     1741 catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc
     1801 catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc
     1861 catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac gtttcccgtt
     1921 gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt tttattgtga
     1981 attctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     2041 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     2101 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     2161 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     2221 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     2281 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     2341 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     2401 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     2461 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     2521 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     2581 gggtttcgcc acctctgact tgagcgtcga tttttgtgat ctcgtcaggg gggcggagcc
     2641 tatggaaaaa cgccagcaac
//
