LOCUS       sequence-495560-        3465 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 3465)
  TITLE     Plant Golden-gate N-Terminal and C-Terminal Tags.
REFERENCE   2  (bases 1 to 3465)
  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: pICSL30061
FEATURES             Location/Qualifiers
     source          1..3465
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             22..1119
                     /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="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1093..1116
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     primer_bind     complement(1306..1323)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1477..2065)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1557..1576)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2236..3027)
                     /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(3249..3353)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3421..3439
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ggtctcacca tggcaaagta taaaatcgaa gaaggtaaac tggtaatctg gattaacggc
       61 gataaaggct ataacggcct cgctgaagtc ggtaagaaat tcgagaaaga taccggaatt
      121 aaagtcaccg ttgagcatcc ggataaactg gaagagaaat tcccacaggt tgcggcaact
      181 ggcgatggcc ctgacattat cttctgggca cacgaccgct ttggtggcta cgctcaatct
      241 ggcctgttgg ctgaaatcac cccggacaaa gcgttccagg acaagctgta tccgtttacc
      301 tgggatgccg tacgttacaa cggcaagctg attgcttacc cgatcgctgt tgaagcgtta
      361 tcgctgattt ataacaaaga tctgctgccg aacccgccaa aaacctggga agagatcccg
      421 gcgctggata aagaactgaa agcgaaaggt aagagcgcgc tgatgttcaa cctgcaagaa
      481 ccgtacttca cctggccgct gattgctgct gacgggggtt atgcgttcaa gtatgaaaac
      541 ggcaagtacg acattaaaga cgtgggcgtg gataacgctg gcgcgaaagc gggtctgacc
      601 ttcctggttg acctgattaa aaacaaacac atgaatgcag acaccgatta ctccatcgca
      661 gaagctgcct ttaataaagg cgaaacagcg atgaccatca acggcccgtg ggcatggtcc
      721 aacatcgaca ccagcaaagt gaattatggt gtaacggtac tgccgacctt caagggtcaa
      781 ccatccaaac cgttcgttgg cgtgctgagc gcaggtatta acgccgccag tccgaacaaa
      841 gagctggcaa aagagttcct cgaaaactat ctgctgactg atgaaggtct ggaagcggtt
      901 aataaagaca aaccgctggg tgccgtagcg ctgaagtctt acgaggaaga gttggcgaaa
      961 gatccacgta ttgccgccac tatggaaaac gcccagaaag gtgaaatcat gccgaacatc
     1021 ccgcagatgt ccgctttctg gtatgccgtg cgtactgcgg tgatcaacgc cgccagcggt
     1081 cgtcagactg tcgatgaagc cctgaaagac gcgcagactg tgaaaattgt gagccgcaaa
     1141 agcctgggcg tgcagaatgt gtatgatatt ggcgtggaaa aagatcataa ctttctgctg
     1201 gcgaatggcc tgattgcgag caacggaatg tgagaccacg agtgattaat gaatcggcca
     1261 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     1321 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     1381 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     1441 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1501 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1561 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1621 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     1681 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1741 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     1801 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1861 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     1921 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1981 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2041 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2101 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2161 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2221 aacttggtct gacagttatt tgccgactac cttggtgatc tcgcctttta cgtaatggac
     2281 aaattcttcc aactgatctg cgcgcgaggc caagcgatct tcttcttgtc caagataagc
     2341 ctgtctagct tcaagtatga cgggctgata ctgggccggc aggcgctcca ttgcccagtc
     2401 ggcagcgaca tccttcggcg cgattttgcc ggttactgcg ctgtaccaaa tgcgggacaa
     2461 cgtaagcact acatttcgct catcgccagc ccagtcgggc ggcgagttcc atagcgttaa
     2521 ggtttcattt agcgcctcaa atagatcctg ttcaggaacc ggatcaaaga gttcctccgc
     2581 cgctggacct accaaggcaa cgctatgttc tcttgctttt gtcagcaaga tagccagatc
     2641 aatgtcgatc gtggctggct cgaagatacc tgcaagaatg tcattgcgct gccattctcc
     2701 aaattgcagt tcgcgcttag ctggataacg ccacggaatg atgtcgtcgt gcacaacaat
     2761 ggtgacttct acagcgcgga gaatctcgct ctctccaggg gaagccgaag tttccaaaag
     2821 gtcgttgatc aaagctcgcc gcgttgtttc atcaagcctt acggtcaccg taaccagcaa
     2881 atcaatatcg ctgtgtggct tcaggccgcc atccactgcg gagccgtaca aatgtacggc
     2941 cagcaacgtc ggttcgagat ggcgctcgat gacgccaact acctctgata gttgagtcga
     3001 tacttcggcg atcaccgctt ccctcatgat gtttaacttt gttttagggc gactgccctg
     3061 ctgcgtaaca tcgttgctgc tccataacat caaacatcga cccacggcgt aacgcgcttg
     3121 ctgcttggat gcccgaggca tagactgtac cccaaaaaaa aacagtcata acaagccatg
     3181 aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt
     3241 gagcgcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     3301 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     3361 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     3421 aataggcgta tcacgaggcc ctttcgtcca ctgaagagcc actgt
//
