LOCUS       sequence-272193-        1969 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 1969)
  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 1969)
  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-G2
FEATURES             Location/Qualifiers
     source          1..1969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     216..238
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(276..294)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(427..1242)
                     /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(427..1242)
                     /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     1150..1169
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1234..1253
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1370..1957
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1859..1878
                     /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 ggcattgcat
      181 ccagccccga gcaaaaggag acccctgact agtctccggg agctgcatgt gtcagaggtt
      241 ttcaccgtca tcaccgaaac gcgcgacacg aaagggcctc gtgatacgcc tatttttata
      301 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggccttttcg gggaaatgtg
      361 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaat
      421 taattcttag aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt
      481 atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca
      541 gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat
      601 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt
      661 gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac
      721 aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg
      781 tgattgcgcc tgagccagac gaaatacgcg gtcgctgtta aaaggacaat tacaaacagg
      841 aatcgaatgc aaccggcgca ggaacaccgc cagcgcatca acaatatttt cacctgaatc
      901 aggatattct tctaatacct ggaatgctgt tttccctggg atcgccgtgg tgagtaacca
      961 tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag
     1021 ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt
     1081 cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg
     1141 cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa
     1201 tcgcggccta gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact
     1261 gtttatgtaa gcagacagtt ttattgtgaa ttctcatgac caaaatccct taacgtgagt
     1321 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     1381 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     1441 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     1501 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     1561 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     1621 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     1681 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     1741 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     1801 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     1861 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     1921 ttttgtgatc tcgtcagggg ggcggagcct atggaaaaac gccagcaac
//
