LOCUS       sequence-456341-        2082 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N 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 2082)
  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 2082)
  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.003
FEATURES             Location/Qualifiers
     source          1..2082
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             17..190
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             191..364
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNGAQAPK"
     CDS             404..424
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      complement(447..481)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(552..1211)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1126..1145
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1393..1981
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1882..1901
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcggtctcaa agaatggtgg acaacaaatt caacaaagaa caacaaaacg cgttctatga
       61 aatcttacat ttacctaact taaacgaaga acaacgaaac gccttcatcc aaagtttaaa
      121 agatgaccca agccaaagcg ctaacctttt agcagaagct aaaaagctaa atgatgctca
      181 ggcgccgaaa gtagacaaca aattcaacaa agaacaacaa aacgcgttct atgaaatctt
      241 acatttacct aacttaaacg aagaacaacg aaacgccttc atccaaagtt taaaagatga
      301 cccaagccaa agcgctaacc ttttagcaga agctaaaaag ctaaatggtg ctcaggcgcc
      361 gaaagagctc aaaaccgcgg ctcttgcgca acacgatgaa gccgagaatt tgtattttca
      421 gggttccggt atgtgagacc agaccaataa aaaacgcccg gcggcaaccg agcgttctga
      481 acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt ccaagcgagc
      541 tcgatatcaa attacgcccc gccctgccac tcatcgcagt actgttgtaa ttcattaagc
      601 attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc
      661 agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc gaagaagttg
      721 tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt ggctgaaacg
      781 aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc gtaacacgcc
      841 acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc actccagagc
      901 gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac actatcccat
      961 atcaccagct caccgtcttt cattgccata cgaaattccg gatgagcatt catcaggcgg
     1021 gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac ggtctttaaa
     1081 aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac tgactgaaat
     1141 gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata tccagtgatt
     1201 tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc
     1261 ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccc gatcaatcat
     1321 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     1381 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     1441 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     1501 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     1561 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     1621 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     1681 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     1741 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     1801 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     1861 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     1921 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     1981 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     2041 gttctttcct gcgttatccc ctgattctgt ggataaccgt ag
//
