LOCUS       sequence-445929-        1945 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  4G-cloning donor for C-terminal TwinStrep tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1945)
  AUTHORS   Taschner M, Dickinson JB, Roisne-Hamelin F, Gruber S
  TITLE     4G cloning: rapid gene assembly for expression of multisubunit 
            protein complexes in diverse hosts
  JOURNAL   bioRxiv 2024.06.17.599261
REFERENCE   2  (bases 1 to 1945)
  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: "bioRxiv 
            2024.06.17.599261"
COMMENT     Sequence Label: pD(Ct)_TS
FEATURES             Location/Qualifiers
     source          1..1945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      196..552
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    complement(605..638)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             722..745
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             782..805
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     promoter        984..1086
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1087..1746
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1153..1172)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ctcaaatgcc tgaggccagt ttgctcaggc tctccccgtg gaggtaataa ttgacgatat
       61 gatcatttat tctgcctccc agctgacatt catccggggt cagcaccgtt tctgcggact
      121 ggctttctac gtgttccgct tcctttagca gcccttgcgc cctgagtgct tgcggcagcg
      181 tgaagctaat tcccatgtca gccgttaagt gttcctgtgt cactcaaaat tgctttgaga
      241 ggctctaagg gcttctcagt gcgttacatc cctggcttgt tgtccacaac cgttaaacct
      301 taaaagcttt aaaagcctta tatattcttt tttttcttat aaaacttaaa accttagagg
      361 ctatttaagt tgctgattta tattaatttt attgttcaaa catgagagct tagtacgtga
      421 aacatgagag cttagtacgt tagccatgag agcttagtac gttagccatg agggtttagt
      481 tcgttaaaca tgagagctta gtacgttaaa catgagagct tagtacgtga aacatgagag
      541 cttagtacgt actatcaaca ggttgaactg ctgatcaaca gatcctctac gcggccgcgg
      601 taccataact tcgtatagca tacattatac gaagttatct gccaggcaca tgggttttac
      661 tagtatcgat tcgcgaccta ctccggaata ttaggtctca ggatctggtg gatctagcgc
      721 ttggagccac ccgcagttcg agaaaggtgg aggttccgga ggtggatcgg gaggtggatc
      781 gtggagccac ccgcagttcg aaaaataagg gagacccact gcttgagcct agaagatccg
      841 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg tcaccgctga gcaataacta
      901 tcataacccc taggtgccat ttcattacct ctttctccgc acccgacata gatctgggcc
      961 aacttttggc gaaaatgaga cgttgatcgg cacgtaagag ggtccaactt tcaccataat
     1021 gaaataagat cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa
     1081 gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt
     1141 aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag
     1201 ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc
     1261 tttattcaca ttcttgcccg cctgatgaat gctcatccgg aattccgtat ggcaatgaaa
     1321 gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa
     1381 actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac
     1441 atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt
     1501 attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta
     1561 aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg
     1621 caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc
     1681 ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggtggg
     1741 gcgtaatttt tttaaggcag ttattggtgc ccttaaacgc ctggttgcta cgcctgaata
     1801 agtgataata agcggatgaa tggcagaaat tcgaaagcaa attcgacccg gtcgtcggtt
     1861 cagggcaggg tcgttaaata gccgcttatg tctattgctg gtttaccggt ttattgacta
     1921 ccggaagcag tgtgaccgtg tgctt
//
