LOCUS       sequence-272109-        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-G3
FEATURES             Location/Qualifiers
     source          1..1972
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     219..241
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(279..297)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(430..1245)
                     /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(430..1245)
                     /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     1153..1172
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1237..1256
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1373..1960
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1862..1881
                     /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 gagggcattg
      181 catccagccc cgagcaaaag gagacccctg actagtctcc gggagctgca tgtgtcagag
      241 gttttcaccg tcatcaccga aacgcgcgac acgaaagggc ctcgtgatac gcctattttt
      301 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggccttt tcggggaaat
      361 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
      421 aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg
      481 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
      541 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
      601 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
      661 agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc
      721 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
      781 tcgtgattgc gcctgagcca gacgaaatac gcggtcgctg ttaaaaggac aattacaaac
      841 aggaatcgaa tgcaaccggc gcaggaacac cgccagcgca tcaacaatat tttcacctga
      901 atcaggatat tcttctaata cctggaatgc tgttttccct gggatcgccg tggtgagtaa
      961 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     1021 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     1081 tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga
     1141 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     1201 taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt
     1261 actgtttatg taagcagaca gttttattgt gaattctcat gaccaaaatc ccttaacgtg
     1321 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     1381 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     1441 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     1501 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     1561 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     1621 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     1681 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     1741 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     1801 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     1861 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     1921 gatttttgtg atctcgtcag gggggcggag cctatggaaa aacgccagca ac
//
