LOCUS       sequence-445941-        2136 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  4G-cloning donor for N-terminal His6-SUMO tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2136)
  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 2136)
  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(Nt)_His6-SUMO
FEATURES             Location/Qualifiers
     source          1..2136
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             41..58
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             68..85
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     promoter        508..610
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             611..1270
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(677..696)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1665..2021
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    complement(2074..2107)
                     /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 tcaggcgagc catcatcatc atcatcacag
       61 cagcggcctg gtgccgcgcg gcagccatat gaatgaccac atcaacctga aggtggccgg
      121 gcaggacggc tccgtggtgc agttcaagat caagaggcac acgccgctga gcaagctgat
      181 gaaggcctac tgcgagaggc agggcttgtc aatgaggcag atcagattca ggttcgacgg
      241 gcagccaatc aatgaaactg acactccagc acagctggag atggaggacg aggacaccat
      301 cgacgtgttc cagcagcaga ccggtggcgc catggagacc cactgcttga gcctagaaga
      361 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgtcaccg ctgagcaata
      421 actatcataa cccctaggtg ccatttcatt acctctttct ccgcacccga catagatctg
      481 ggccaacttt tggcgaaaat gagacgttga tcggcacgta agagggtcca actttcacca
      541 taatgaaata agatcactac cgggcgtatt ttttgagtta tcgagatttt caggagctaa
      601 ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca
      661 tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt
      721 tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc
      781 ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaattcc gtatggcaat
      841 gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga
      901 gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct
      961 acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg
     1021 gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga
     1081 tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta
     1141 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga
     1201 tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg
     1261 tggggcgtaa tttttttaag gcagttattg gtgcccttaa acgcctggtt gctacgcctg
     1321 aataagtgat aataagcgga tgaatggcag aaattcgaaa gcaaattcga cccggtcgtc
     1381 ggttcagggc agggtcgtta aatagccgct tatgtctatt gctggtttac cggtttattg
     1441 actaccggaa gcagtgtgac cgtgtgcttc tcaaatgcct gaggccagtt tgctcaggct
     1501 ctccccgtgg aggtaataat tgacgatatg atcatttatt ctgcctccca gctgacattc
     1561 atccggggtc agcaccgttt ctgcggactg gctttctacg tgttccgctt cctttagcag
     1621 cccttgcgcc ctgagtgctt gcggcagcgt gaagctaatt cccatgtcag ccgttaagtg
     1681 ttcctgtgtc actcaaaatt gctttgagag gctctaaggg cttctcagtg cgttacatcc
     1741 ctggcttgtt gtccacaacc gttaaacctt aaaagcttta aaagccttat atattctttt
     1801 ttttcttata aaacttaaaa ccttagaggc tatttaagtt gctgatttat attaatttta
     1861 ttgttcaaac atgagagctt agtacgtgaa acatgagagc ttagtacgtt agccatgaga
     1921 gcttagtacg ttagccatga gggtttagtt cgttaaacat gagagcttag tacgttaaac
     1981 atgagagctt agtacgtgaa acatgagagc ttagtacgta ctatcaacag gttgaactgc
     2041 tgatcaacag atcctctacg cggccgcggt accataactt cgtatagcat acattatacg
     2101 aagttatctg ccaggcacat gggttttact agtatc
//
