LOCUS       sequence-272206-        1987 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 1987)
  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 1987)
  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-GGASPAGG
FEATURES             Location/Qualifiers
     source          1..1987
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     234..256
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(294..312)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(445..1260)
                     /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(445..1260)
                     /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     1168..1187
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1252..1271
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1388..1975
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1877..1896
                     /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 gagcatctcc
      181 tgcaggaggg cattgcatcc agccccgagc aaaaggagac ccctgactag tctccgggag
      241 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgacacgaa agggcctcgt
      301 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
      361 ccttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
      421 atgtatccgc tcatgaatta attcttagaa aaactcatcg agcatcaaat gaaactgcaa
      481 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
      541 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
      601 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
      661 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc
      721 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac
      781 caaaccgtta ttcattcgtg attgcgcctg agccagacga aatacgcggt cgctgttaaa
      841 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacaccgcca gcgcatcaac
      901 aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt tccctgggat
      961 cgccgtggtg agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag
     1021 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac
     1081 gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata
     1141 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc
     1201 atccatgttg gaatttaatc gcggcctaga gcaagacgtt tcccgttgaa tatggctcat
     1261 aacacccctt gtattactgt ttatgtaagc agacagtttt attgtgaatt ctcatgacca
     1321 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     1381 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     1441 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     1501 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     1561 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     1621 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     1681 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     1741 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     1801 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     1861 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     1921 tctgacttga gcgtcgattt ttgtgatctc gtcagggggg cggagcctat ggaaaaacgc
     1981 cagcaac
//
