LOCUS       sequence-495490-        2526 bp    DNA     circular SYN 17-DEC-2025
DEFINITION  Level 0 Golden Gate part N-Terminal Tag with overhangs CCAT - AATG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2526)
  TITLE     Plant Golden-gate N-Terminal and C-Terminal Tags.
REFERENCE   2  (bases 1 to 2526)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pICSL30063
FEATURES             Location/Qualifiers
     source          1..2526
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             22..39
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             67..87
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             94..159
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(367..384)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(538..1126)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(618..637)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1297..2088)
                     /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"
     promoter        complement(2310..2414)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2482..2500
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ggtctcacca tggcaaagta tcatcaccat caccatcacg attatgatat tccaactact
       61 gctagcgaga atttgtattt tcagggtgag ctcgactaca aagaccatga cggtgattat
      121 aaagatcatg acatcgacta caaggatgac gatgacaaga acgcggctat tagatcagga
      181 gtgaaaattg tgagccgcaa aagcctgggc gtgcagaatg tgtatgatat tggcgtggaa
      241 aaagatcata actttctgct ggcgaatggc ctgattgcga gcaacggaat gtgagaccac
      301 gagtgattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
      361 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
      421 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
      481 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
      541 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
      601 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
      661 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
      721 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
      781 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
      841 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
      901 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
      961 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     1021 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     1081 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     1141 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     1201 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     1261 aatctaaagt atatatgagt aaacttggtc tgacagttat ttgccgacta ccttggtgat
     1321 ctcgcctttt acgtaatgga caaattcttc caactgatct gcgcgcgagg ccaagcgatc
     1381 ttcttcttgt ccaagataag cctgtctagc ttcaagtatg acgggctgat actgggccgg
     1441 caggcgctcc attgcccagt cggcagcgac atccttcggc gcgattttgc cggttactgc
     1501 gctgtaccaa atgcgggaca acgtaagcac tacatttcgc tcatcgccag cccagtcggg
     1561 cggcgagttc catagcgtta aggtttcatt tagcgcctca aatagatcct gttcaggaac
     1621 cggatcaaag agttcctccg ccgctggacc taccaaggca acgctatgtt ctcttgcttt
     1681 tgtcagcaag atagccagat caatgtcgat cgtggctggc tcgaagatac ctgcaagaat
     1741 gtcattgcgc tgccattctc caaattgcag ttcgcgctta gctggataac gccacggaat
     1801 gatgtcgtcg tgcacaacaa tggtgacttc tacagcgcgg agaatctcgc tctctccagg
     1861 ggaagccgaa gtttccaaaa ggtcgttgat caaagctcgc cgcgttgttt catcaagcct
     1921 tacggtcacc gtaaccagca aatcaatatc gctgtgtggc ttcaggccgc catccactgc
     1981 ggagccgtac aaatgtacgg ccagcaacgt cggttcgaga tggcgctcga tgacgccaac
     2041 tacctctgat agttgagtcg atacttcggc gatcaccgct tccctcatga tgtttaactt
     2101 tgttttaggg cgactgccct gctgcgtaac atcgttgctg ctccataaca tcaaacatcg
     2161 acccacggcg taacgcgctt gctgcttgga tgcccgaggc atagactgta ccccaaaaaa
     2221 aaacagtcat aacaagccat gaaaaccgcc actgcgccgt taccaccgct gcgttcggtc
     2281 aaggttctgg accagttgcg tgagcgcata ctcttccttt ttcaatatta ttgaagcatt
     2341 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     2401 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     2461 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtcc actgaagagc
     2521 cactgt
//
