LOCUS       sequence-272208-        2270 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 2270)
  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 2270)
  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: pFD-FN3
FEATURES             Location/Qualifiers
     source          1..2270
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     517..539
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(577..595)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(728..1543)
                     /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(728..1543)
                     /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     1451..1470
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1535..1554
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1671..2258
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2160..2179
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
       61 ttatcccctg attctgtgga taaccgtatt accgccaatt gagtgagctg ataccgctcg
      121 ccgcagccga acgaccgagc gcagcgagtc acccgcgaaa tacaagaaga caattttccc
      181 gcaccagcag tgggggggtt agttcggtgc ctacaaattt ggaggtggtt gcggcaacac
      241 ccacctcctt gctgatctct tgggacgcct accgtgagtt acccgtgtcc tactaccgta
      301 tcacgtacgg tgagactggc gggaactcac ctgtgcagga atttacagtg ccggggtcca
      361 aatcaaccgc taccatctcg ggattgaagc cgggtgttga ttatactatc actgtgtatg
      421 cccactataa ctaccactac tatagttccc ccatctccat caactatcgc ggcattgcat
      481 aaagccccga cagtaaaaag agaccctgac tagtctccgg gagctgcatg tgtcagaggt
      541 tttcaccgtc atcaccgaaa cgcgcgacac gaaagggcct cgtgatacgc ctatttttat
      601 aggttaatgt catgataata atggtttctt agacgtcagg tggccttttc ggggaaatgt
      661 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgaa
      721 ttaattctta gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat
      781 tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc
      841 agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa
      901 tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag
      961 tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag acttgttcaa
     1021 caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc
     1081 gtgattgcgc ctgagccaga cgaaatacgc ggtcgctgtt aaaaggacaa ttacaaacag
     1141 gaatcgaatg caaccggcgc aggaacaccg ccagcgcatc aacaatattt tcacctgaat
     1201 caggatattc ttctaatacc tggaatgctg ttttccctgg gatcgccgtg gtgagtaacc
     1261 atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca
     1321 gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt
     1381 tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt
     1441 gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta
     1501 atcgcggcct agagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac
     1561 tgtttatgta agcagacagt tttattgtga attctcatga ccaaaatccc ttaacgtgag
     1621 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     1681 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     1741 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     1801 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     1861 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     1921 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     1981 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2041 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     2101 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     2161 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     2221 tttttgtgat ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
//
