LOCUS       sequence-272215-        1990 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 1990)
  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 1990)
  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)7G
FEATURES             Location/Qualifiers
     source          1..1990
                     /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     967..989
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1027..1045)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(join(1178..1990,1..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(1178..1990,1..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     1901..1920
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     join(1985..1990,1..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 gggcattgca tccagccccg agcaaaagga gacccctgac
      961 tagtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgacac
     1021 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     1081 agacgtcagg tggccttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     1141 aatacattca aatatgtatc cgctcatgaa ttaattctta gaaaaactca tcgagcatca
     1201 aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt
     1261 tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc
     1321 ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa
     1381 taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa
     1441 gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat
     1501 cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagccaga cgaaatacgc
     1561 ggtcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacaccg
     1621 ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg
     1681 ttttccctgg gatcgccgtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct
     1741 tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa
     1801 catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc
     1861 catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc
     1921 catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac gtttcccgtt
     1981 gaatatggct
//
