LOCUS       sequence-296181-        2126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein tags.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2126)
  AUTHORS   Jaramillo-Madrid AC, Abbriano R, Ashworth J, Fabris M, Pernice M, 
            Ralph PJ
  TITLE     Overexpression of Key Sterol Pathway Enzymes in Two Model Marine 
            Diatoms Alters Sterol Profiles in Phaeodactylum tricornutum.
  JOURNAL   Pharmaceuticals (Basel). 2020 Dec 21;13(12). pii: ph13120481. doi: 
            10.3390/ph13120481.
  PUBMED    33371196
REFERENCE   2  (bases 1 to 2126)
  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: 
            "Pharmaceuticals (Basel)."; date: "2020-12-21"; volume: "13(12). 
            pii: ph13120481. doi"; pages: " 10.3390/ph13120481"
COMMENT     Sequence Label: DE_Thr6XHisFLAG
FEATURES             Location/Qualifiers
     source          1..2126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             431..448
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             449..466
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             467..490
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     rep_origin      complement(636..1224)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(716..735)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1315..2106)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 tctcttgatc agatattgat cccctgcgcc atcagatcct tggcggcaag aaagccatcc
       61 agtttacttt gcagggcttc ccaaccttac cagagggcgc cccagctggc aattccggtt
      121 cgcttgctaa gcttgcatgc ctgcaagtcg actctagagg agcggtgatc acaggcagca
      181 acgctctgtc atcgttacaa tcaacatgct accctccgcg agatcatccg tgtttcaaac
      241 ccggcagctt agttgccgtt cttccgaata gcatcggtaa catgagcaaa gtctgccgcc
      301 ttacaacggc tctcccgctg acgccgtccc ggactgatgg gctgcctgta tcgagtggtg
      361 attttgtgcc gagctcatta ctcgcatcca ttctcaggct gtctcgtctc gtctcaggtc
      421 tccaggtgcg ctggtccctc gcggtagcca ccaccaccat caccacgact acaaggatga
      481 tgacgataag tgagcttcga gaccaccagg atacatagat taccacaact ccgagccctt
      541 ccacccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
      601 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
      661 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
      721 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
      781 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
      841 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
      901 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
      961 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     1021 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     1081 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     1141 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     1201 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     1261 cagtggaacg aaaactcacg ttaagggatt ttggtcatgc attctaggtg attattattt
     1321 gccgactacc ttggtgatct cgcctttcac gtagtggaca aattcttcca actgatctgc
     1381 gcgcgaggcc aagcgatctt cttcttgtcc aagataagcc tgtctagctt caagtatgac
     1441 gggctgatac tgggccggca ggcgctccat tgcccagtcg gcagcgacat ccttcggcgc
     1501 gattttgccg gttactgcgc tgtaccaaat gcgggacaac gtaagcacta catttcgctc
     1561 atcgccagcc cagtcgggcg gcgagttcca tagcgttaag gtttcattta gcgcctcaaa
     1621 tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc gctggaccta ccaaggcaac
     1681 gctatgttct cttgcttttg tcagcaagat agccagatca atgtcgatcg tggctggctc
     1741 gaagatacca gcaagaatgt cattgcgctg ccattctcca aattgcagtt cgcgcttagc
     1801 tggataacgc cacggaatga tgtcgtcgtg cacaacaatg gtgacttcta cagcgcggag
     1861 aatctcactc tctccagggg aagccgaagt ttccaaaagg tcgttgatca aagctcgccg
     1921 cgttgtttca tcaagcctta cggtcaccgt aaccagcaaa tcaatatcac tgtgtggctt
     1981 caggccgcca tccactgcgg agccgtacaa atgtacggcc agcaacgtcg gttcgagatg
     2041 gcgctcgatg acgccaacta cctctgatag ttgagtcgat acttcggcga tcaccgcttc
     2101 cctcatgcga aacgatcctc atcctg
//
