LOCUS       sequence-272103-        1972 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 1972)
  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 1972)
  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-GPG
FEATURES             Location/Qualifiers
     source          1..1972
                     /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     949..971
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1009..1027)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(join(1160..1972,1..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(1160..1972,1..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     1883..1902
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     join(1967..1972,1..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 cggggcattg catccagccc cgagcaaaag gagacccctg actagtctcc gggagctgca
      961 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgac acgaaagggc ctcgtgatac
     1021 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggccttt
     1081 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     1141 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat
     1201 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
     1261 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
     1321 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
     1381 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
     1441 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
     1501 cgttattcat tcgtgattgc gcctgagcca gacgaaatac gcggtcgctg ttaaaaggac
     1561 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac cgccagcgca tcaacaatat
     1621 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttccct gggatcgccg
     1681 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     1741 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     1801 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
     1861 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     1921 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ct
//
