LOCUS       sequence-495567-        3468 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 3468)
  TITLE     Plant Golden-gate N-Terminal and C-Terminal Tags.
REFERENCE   2  (bases 1 to 3468)
  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: pICSL30071
FEATURES             Location/Qualifiers
     source          1..3468
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             16..33
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             97..1197
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1171..1194
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1207..1230
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(1309..1326)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1480..2068)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1560..1579)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2239..3030)
                     /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(3252..3356)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3424..3442
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ggtctcacca tggcacacca tcaccaccat cacagcagcg gtggactgaa tgatatattc
       61 gaggctcaga agattgaata ccacgagagc agcggcatga aaatcgaaga aggtaaactg
      121 gtaatctgga ttaacggcga taaaggctat aacggcctcg ctgaagtcgg taagaaattc
      181 gagaaagata ccggaattaa agtcaccgtt gagcatccgg ataaactgga agagaaattc
      241 ccacaggttg cggcaactgg cgatggccct gacattatct tctgggcaca cgaccgcttt
      301 ggtggctacg ctcaatctgg cctgttggct gaaatcaccc cggacaaagc gttccaggac
      361 aagctgtatc cgtttacctg ggatgccgta cgttacaacg gcaagctgat tgcttacccg
      421 atcgctgttg aagcgttatc gctgatttat aacaaagatc tgctgccgaa cccgccaaaa
      481 acctgggaag agatcccggc gctggataaa gaactgaaag cgaaaggtaa gagcgcgctg
      541 atgttcaacc tgcaagaacc gtacttcacc tggccgctga ttgctgctga cgggggttat
      601 gcgttcaagt atgaaaacgg caagtacgac attaaagacg tgggcgtgga taacgctggc
      661 gcgaaagcgg gtctgacctt cctggttgac ctgattaaaa acaaacacat gaatgcagac
      721 accgattact ccatcgcaga agctgccttt aataaaggcg aaacagcgat gaccatcaac
      781 ggcccgtggg catggtccaa catcgacacc agcaaagtga attatggtgt aacggtactg
      841 ccgaccttca agggtcaacc atccaaaccg ttcgttggcg tgctgagcgc aggtattaac
      901 gccgccagtc cgaacaaaga gctggcaaaa gagttcctcg aaaactatct gctgactgat
      961 gaaggtctgg aagcggttaa taaagacaaa ccgctgggtg ccgtagcgct gaagtcttac
     1021 gaggaagagt tggcgaaaga tccacgtatt gccgccacta tggaaaacgc ccagaaaggt
     1081 gaaatcatgc cgaacatccc gcagatgtcc gctttctggt atgccgtgcg tactgcggtg
     1141 atcaacgccg ccagcggtcg tcagactgtc gatgaagccc tgaaagacgc gcagactagc
     1201 agcggtctgg aagttctgtt tcagggccca atgtgagacc acgagtgatt aatgaatcgg
     1261 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     1321 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     1381 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     1441 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1501 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1561 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1621 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1681 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1741 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1801 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1861 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1921 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1981 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2041 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2101 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2161 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2221 gtaaacttgg tctgacagtt atttgccgac taccttggtg atctcgcctt ttacgtaatg
     2281 gacaaattct tccaactgat ctgcgcgcga ggccaagcga tcttcttctt gtccaagata
     2341 agcctgtcta gcttcaagta tgacgggctg atactgggcc ggcaggcgct ccattgccca
     2401 gtcggcagcg acatccttcg gcgcgatttt gccggttact gcgctgtacc aaatgcggga
     2461 caacgtaagc actacatttc gctcatcgcc agcccagtcg ggcggcgagt tccatagcgt
     2521 taaggtttca tttagcgcct caaatagatc ctgttcagga accggatcaa agagttcctc
     2581 cgccgctgga cctaccaagg caacgctatg ttctcttgct tttgtcagca agatagccag
     2641 atcaatgtcg atcgtggctg gctcgaagat acctgcaaga atgtcattgc gctgccattc
     2701 tccaaattgc agttcgcgct tagctggata acgccacgga atgatgtcgt cgtgcacaac
     2761 aatggtgact tctacagcgc ggagaatctc gctctctcca ggggaagccg aagtttccaa
     2821 aaggtcgttg atcaaagctc gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag
     2881 caaatcaata tcgctgtgtg gcttcaggcc gccatccact gcggagccgt acaaatgtac
     2941 ggccagcaac gtcggttcga gatggcgctc gatgacgcca actacctctg atagttgagt
     3001 cgatacttcg gcgatcaccg cttccctcat gatgtttaac tttgttttag ggcgactgcc
     3061 ctgctgcgta acatcgttgc tgctccataa catcaaacat cgacccacgg cgtaacgcgc
     3121 ttgctgcttg gatgcccgag gcatagactg taccccaaaa aaaaacagtc ataacaagcc
     3181 atgaaaaccg ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct ggaccagttg
     3241 cgtgagcgca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     3301 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     3361 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     3421 aaaaataggc gtatcacgag gccctttcgt ccactgaaga gccactgt
//
