LOCUS       sequence-456354-        2089 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C terminal dual protein A affinity tag with TEV cleavage site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2089)
  AUTHORS   Lai Z, Flanigan SF, Boudes M, Davidovich C
  TITLE     Modular cloning of multigene vectors for the baculovirus system and 
            yeast.
  JOURNAL   J Mol Biol. 2025 Jan 13:168943. doi: 10.1016/j.jmb.2025.168943.
  PUBMED    39814169
REFERENCE   2  (bases 1 to 2089)
  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: "J Mol Biol.
            2025 Jan 13:168943. doi: 10.1016/j.jmb.2025.168943."
COMMENT     Sequence Label: pM.004
FEATURES             Location/Qualifiers
     source          1..2089
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             20..40
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             80..253
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             254..427
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNGAQAPK"
     terminator      complement(454..488)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(559..1218)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1133..1152
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1400..1988
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1889..1908
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcggtctcaa tcctcaggtg agaatttgta ttttcagggt gagctcaaaa ccgcggctct
       61 tgcgcaacac gatgaagccg tggacaacaa attcaacaaa gaacaacaaa acgcgttcta
      121 tgaaatctta catttaccta acttaaacga agaacaacga aacgccttca tccaaagttt
      181 aaaagatgac ccaagccaaa gcgctaacct tttagcagaa gctaaaaagc taaatgatgc
      241 tcaggcgccg aaagtagaca acaaattcaa caaagaacaa caaaacgcgt tctatgaaat
      301 cttacattta cctaacttaa acgaagaaca acgaaacgcc ttcatccaaa gtttaaaaga
      361 tgacccaagc caaagcgcta accttttagc agaagctaaa aagctaaatg gtgctcaggc
      421 gccgaaataa ctcgagtggc tgagaccaga ccaataaaaa acgcccggcg gcaaccgagc
      481 gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca
      541 agcgagctcg atatcaaatt acgccccgcc ctgccactca tcgcagtact gttgtaattc
      601 attaagcatt ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag
      661 cggcatcagc accttgtcgc cttgcgtata atatttgccc atggtgaaaa cgggggcgaa
      721 gaagttgtcc atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc
      781 tgaaacgaaa aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta
      841 acacgccaca tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact
      901 ccagagcgat gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact
      961 atcccatatc accagctcac cgtctttcat tgccatacga aattccggat gagcattcat
     1021 caggcgggca agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt
     1081 ctttaaaaag gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga
     1141 ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc
     1201 agtgattttt ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa
     1261 tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgcccgat
     1321 caatcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1381 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1441 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1501 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     1561 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1621 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1681 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1741 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1801 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1861 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1921 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1981 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     2041 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtag
//
