LOCUS       sequence-445934-        1894 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  4G-cloning donor for C-terminal 3C-His8 tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1894)
  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 1894)
  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)_3C-His8
FEATURES             Location/Qualifiers
     source          1..1894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             40..63
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             64..87
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     promoter        266..368
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             369..1028
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(435..454)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1423..1779
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    complement(1832..1865)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 gattcgcgac ctactccgga atattaggtc tcaggatccc tggaagttct gttccagggg
       61 ccccaccatc accatcacca tcaccattaa gggagaccca ctgcttgagc ctagaagatc
      121 cggctgctaa caaagcccga aaggaagctg agttggctgc tgtcaccgct gagcaataac
      181 tatcataacc cctaggtgcc atttcattac ctctttctcc gcacccgaca tagatctggg
      241 ccaacttttg gcgaaaatga gacgttgatc ggcacgtaag agggtccaac tttcaccata
      301 atgaaataag atcactaccg ggcgtatttt ttgagttatc gagattttca ggagctaagg
      361 aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc
      421 gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc
      481 agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg
      541 cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattccgt atggcaatga
      601 aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc
      661 aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac
      721 acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt
      781 ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt
      841 taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata
      901 cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg
      961 gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggtg
     1021 gggcgtaatt tttttaaggc agttattggt gcccttaaac gcctggttgc tacgcctgaa
     1081 taagtgataa taagcggatg aatggcagaa attcgaaagc aaattcgacc cggtcgtcgg
     1141 ttcagggcag ggtcgttaaa tagccgctta tgtctattgc tggtttaccg gtttattgac
     1201 taccggaagc agtgtgaccg tgtgcttctc aaatgcctga ggccagtttg ctcaggctct
     1261 ccccgtggag gtaataattg acgatatgat catttattct gcctcccagc tgacattcat
     1321 ccggggtcag caccgtttct gcggactggc tttctacgtg ttccgcttcc tttagcagcc
     1381 cttgcgccct gagtgcttgc ggcagcgtga agctaattcc catgtcagcc gttaagtgtt
     1441 cctgtgtcac tcaaaattgc tttgagaggc tctaagggct tctcagtgcg ttacatccct
     1501 ggcttgttgt ccacaaccgt taaaccttaa aagctttaaa agccttatat attctttttt
     1561 ttcttataaa acttaaaacc ttagaggcta tttaagttgc tgatttatat taattttatt
     1621 gttcaaacat gagagcttag tacgtgaaac atgagagctt agtacgttag ccatgagagc
     1681 ttagtacgtt agccatgagg gtttagttcg ttaaacatga gagcttagta cgttaaacat
     1741 gagagcttag tacgtgaaac atgagagctt agtacgtact atcaacaggt tgaactgctg
     1801 atcaacagat cctctacgcg gccgcggtac cataacttcg tatagcatac attatacgaa
     1861 gttatctgcc aggcacatgg gttttactag tatc
//
