LOCUS       sequence-272110-        2011 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 2011)
  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 2011)
  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-GGA(EAAAK)2AGG
FEATURES             Location/Qualifiers
     source          1..2011
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     258..280
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(318..336)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(469..1284)
                     /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(469..1284)
                     /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     1192..1211
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1276..1295
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1412..1999
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1901..1920
                     /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 gagcagaagc
      181 ggccgctaaa gaggcagctg caaaagcagg agggcattgc atccagcccc gagcaaaagg
      241 agacccctga ctagtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
      301 acgcgcgaca cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
      361 aatggtttct tagacgtcag gtggcctttt cggggaaatg tgcgcggaac ccctatttgt
      421 ttatttttct aaatacattc aaatatgtat ccgctcatga attaattctt agaaaaactc
      481 atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg
      541 aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag
      601 atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc
      661 ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga
      721 gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc
      781 gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagccag
      841 acgaaatacg cggtcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg
      901 caggaacacc gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac
      961 ctggaatgct gttttccctg ggatcgccgt ggtgagtaac catgcatcat caggagtacg
     1021 gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat
     1081 ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc
     1141 atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc
     1201 ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc tagagcaaga
     1261 cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag
     1321 ttttattgtg aattctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1381 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1441 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1501 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1561 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1621 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1681 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1741 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1801 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1861 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1921 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tctcgtcagg
     1981 ggggcggagc ctatggaaaa acgccagcaa c
//
