LOCUS       sequence-445967-        2020 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  4G-cloning donor for C-terminal AviTag-3C-TwinStrep tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2020)
  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 2020)
  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)_Avi-3C-TS
FEATURES             Location/Qualifiers
     source          1..2020
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             46..90
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             91..114
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             130..153
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             190..213
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     promoter        392..494
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             495..1154
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(561..580)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1549..1905
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    complement(1958..1991)
                     /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 tcaggatccg caggcggcct gaatgatatt
       61 tttgaagcac aaaaaattga atggcatgaa ctggaagttc tgttccaggg gcccggtgga
      121 tctagcgctt ggagccaccc gcagttcgag aaaggtggag gttccggagg tggatcggga
      181 ggtggatcgt ggagccaccc gcagttcgaa aaataaggga gacccactgc ttgagcctag
      241 aagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgtc accgctgagc
      301 aataactatc ataaccccta ggtgccattt cattacctct ttctccgcac ccgacataga
      361 tctgggccaa cttttggcga aaatgagacg ttgatcggca cgtaagaggg tccaactttc
      421 accataatga aataagatca ctaccgggcg tattttttga gttatcgaga ttttcaggag
      481 ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat
      541 ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga
      601 ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt
      661 atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg
      721 caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc
      781 atgagcaaac tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt
      841 ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta
      901 aagggtttat tgagaatatg tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt
      961 ttgatttaaa cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat
     1021 attatacgca aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgttt
     1081 gtgatggctt ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc
     1141 agggtggggc gtaatttttt taaggcagtt attggtgccc ttaaacgcct ggttgctacg
     1201 cctgaataag tgataataag cggatgaatg gcagaaattc gaaagcaaat tcgacccggt
     1261 cgtcggttca gggcagggtc gttaaatagc cgcttatgtc tattgctggt ttaccggttt
     1321 attgactacc ggaagcagtg tgaccgtgtg cttctcaaat gcctgaggcc agtttgctca
     1381 ggctctcccc gtggaggtaa taattgacga tatgatcatt tattctgcct cccagctgac
     1441 attcatccgg ggtcagcacc gtttctgcgg actggctttc tacgtgttcc gcttccttta
     1501 gcagcccttg cgccctgagt gcttgcggca gcgtgaagct aattcccatg tcagccgtta
     1561 agtgttcctg tgtcactcaa aattgctttg agaggctcta agggcttctc agtgcgttac
     1621 atccctggct tgttgtccac aaccgttaaa ccttaaaagc tttaaaagcc ttatatattc
     1681 ttttttttct tataaaactt aaaaccttag aggctattta agttgctgat ttatattaat
     1741 tttattgttc aaacatgaga gcttagtacg tgaaacatga gagcttagta cgttagccat
     1801 gagagcttag tacgttagcc atgagggttt agttcgttaa acatgagagc ttagtacgtt
     1861 aaacatgaga gcttagtacg tgaaacatga gagcttagta cgtactatca acaggttgaa
     1921 ctgctgatca acagatcctc tacgcggccg cggtaccata acttcgtata gcatacatta
     1981 tacgaagtta tctgccaggc acatgggttt tactagtatc
//
