LOCUS       sequence-445932-        1870 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  4G-cloning donor for C-terminal His8 tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1870)
  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 1870)
  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)_His8
FEATURES             Location/Qualifiers
     source          1..1870
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             40..63
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     promoter        242..344
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             345..1004
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(411..430)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1399..1755
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    complement(1808..1841)
                     /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 accatcacca tcaccatcac
       61 cattaaggga gacccactgc ttgagcctag aagatccggc tgctaacaaa gcccgaaagg
      121 aagctgagtt ggctgctgtc accgctgagc aataactatc ataaccccta ggtgccattt
      181 cattacctct ttctccgcac ccgacataga tctgggccaa cttttggcga aaatgagacg
      241 ttgatcggca cgtaagaggg tccaactttc accataatga aataagatca ctaccgggcg
      301 tattttttga gttatcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca
      361 ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc
      421 agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa
      481 agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc
      541 tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga cggtgagctg gtgatatggg
      601 atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct
      661 ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt
      721 gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct
      781 cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact
      841 tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc
      901 cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta
      961 atgaattaca acagtactgc gatgagtggc agggtggggc gtaatttttt taaggcagtt
     1021 attggtgccc ttaaacgcct ggttgctacg cctgaataag tgataataag cggatgaatg
     1081 gcagaaattc gaaagcaaat tcgacccggt cgtcggttca gggcagggtc gttaaatagc
     1141 cgcttatgtc tattgctggt ttaccggttt attgactacc ggaagcagtg tgaccgtgtg
     1201 cttctcaaat gcctgaggcc agtttgctca ggctctcccc gtggaggtaa taattgacga
     1261 tatgatcatt tattctgcct cccagctgac attcatccgg ggtcagcacc gtttctgcgg
     1321 actggctttc tacgtgttcc gcttccttta gcagcccttg cgccctgagt gcttgcggca
     1381 gcgtgaagct aattcccatg tcagccgtta agtgttcctg tgtcactcaa aattgctttg
     1441 agaggctcta agggcttctc agtgcgttac atccctggct tgttgtccac aaccgttaaa
     1501 ccttaaaagc tttaaaagcc ttatatattc ttttttttct tataaaactt aaaaccttag
     1561 aggctattta agttgctgat ttatattaat tttattgttc aaacatgaga gcttagtacg
     1621 tgaaacatga gagcttagta cgttagccat gagagcttag tacgttagcc atgagggttt
     1681 agttcgttaa acatgagagc ttagtacgtt aaacatgaga gcttagtacg tgaaacatga
     1741 gagcttagta cgtactatca acaggttgaa ctgctgatca acagatcctc tacgcggccg
     1801 cggtaccata acttcgtata gcatacatta tacgaagtta tctgccaggc acatgggttt
     1861 tactagtatc
//
