LOCUS       sequence-271257-        3331 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 3331)
  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 3331)
  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: pFLinkC-XE
FEATURES             Location/Qualifiers
     source          1..3331
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     60..79
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        60..78
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(1206..1252)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1290..1317
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     1326..1348
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1361..1379)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1453..1557
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1558..1626,1627..2418)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1776..1795)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2584..3171
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3073..3092
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa attctagagt
       61 aatacgactc actataggga gaaataattt tgtttaactt taaaaaaaaa aagaaggaga
      121 ataatctatg gggcactgca tgggggtgtc caagggagag gaggataata tggcttccct
      181 gccggcgacc catgaacttc atatctttgg ttcgatcaac ggagtggact ttgatatggt
      241 tggtcaaggc acgggaaacc cgaacgatgg gtatgaagag ttgaatctta aatcgacaaa
      301 aggggatctt caattcagcc cttggattct ggtcccgcat atcgggtatg gcttccacca
      361 gtatttacca tacccggacg ggatgtctcc attccaggcc gctatggtgg atggctctgg
      421 ttaccaagtg catcgcacca tgcagttcga ggacggagca tctcttactg ttaactaccg
      481 ttatacctac gaaggatcac acattaaggg agaagctcag gttaaaggaa cgggatttcc
      541 cgctgacgga cccgtaatga ctaatagcct gacagctgcc gactggtgtc gcagtaagaa
      601 aacttatcct aacgacaaaa cgattatctc gacgttcaaa tggtcttaca cgactggaaa
      661 tgggaagcgc taccgtagca cggcccgcac gacgtacacc tttgccaaac cgatggcggc
      721 taactatctg aagaatcaac ccatgtatgt ttttcgtaaa acagaattga aacatagtaa
      781 gactgaactg aacttcaagg agtggcaaaa agcatttacg gacgtgatgg ggatggatga
      841 gttatataaa ggtggatccg gaggtagctg tctgagctac gacactgaaa tcctgacggt
      901 agaatacggg ttccttccca tcggtaagat cgttgaagaa cgtatcgagt gtactgttta
      961 cactgtggac aaaaacgggt tcgtctatac gcagccgatt gcacaatggc ataatcgcgg
     1021 cgaacaagag gtttttgagt attgcctgga ggacggcagt atcattcgcg ccactaaaga
     1081 tcataaattt atgacgaccg atggtcagat gttgcctatt gatgaaatct tcgaacgtgg
     1141 gcttgacttg aaacaagttg atggcttacc aggcattgca taagctagcc gattccaggc
     1201 atcaaataaa acgaaaggcc cagtctttcg actgagcctt tcgttttatc tgttgtttgt
     1261 cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt
     1321 atactccggg agctgcatgt gtcagaggtt taacgaaagg gcctcgtgat acgcctattt
     1381 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcag gcttactttt
     1441 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     1501 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     1561 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     1621 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     1681 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     1741 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     1801 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     1861 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgt
     1921 agtgctgcca taaccatgag tgataacaca gcggccaact tacttctgac aacgatcgga
     1981 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     2041 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     2101 gtagcgatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     2161 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tcttcgctca
     2221 gcacttccag ctggttggtt tattgctgat aaatctggag ccggtgagcg tggatctcgc
     2281 ggtatcatag cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     2341 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     2401 ctgattaagc attggtaact gtcagaccaa gtttactcat atatactttg aattcaaact
     2461 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     2521 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2581 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2641 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2701 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     2761 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2821 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2881 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2941 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3001 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3061 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3121 acttgagcgt cgatttttgt gatctcgtca ggggggcgga gcctatggaa aaacgccagc
     3181 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3241 gcgttatccc ctgattctgt ggataaccgt attaccgcca attgatgtcg gcgatatagg
     3301 cgccagcaac cgcacctgtg gcgccggtga t
//
