LOCUS       sequence-495472-        3384 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 3384)
  TITLE     Plant Golden-gate N-Terminal and C-Terminal Tags.
REFERENCE   2  (bases 1 to 3384)
  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: pICSL30034
FEATURES             Location/Qualifiers
     source          1..3384
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             70..93
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             130..1089
                     /codon_start=1
                     /product="mutant of the E. coli biotin ligase BioID 
                     engineered for improved catalytic efficiency (Branon et 
                     al., 2018)"
                     /label=TurboID
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     primer_bind     complement(1225..1242)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1396..1984)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1476..1495)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2155..2946)
                     /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(3168..3272)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3340..3358
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ggtctcacca tggattataa ggaccatgac ggagactata aggaccatga cctcgacgct
       61 gcagcagctg attataagga cgatgacgat aaaggcggag gttcaggggg tggctcaggt
      121 ggcggatcaa tgaaagacaa tactgtgcct ctgaagctga tcgctctcct ggctaatggc
      181 gagttccata gtggcgaaca gctgggagaa accctgggca tgtccagggc cgctatcaac
      241 aagcacattc agactctgcg cgactggggc gtggacgtgt tcaccgtgcc cggaaagggc
      301 tactctctgc ccgagcctat cccgctgctg aacgctaaac agattttggg acagctggac
      361 ggcgggagcg tggcagtcct gcctgtggtc gactccacca atcagtacct gctggatcga
      421 atcggcgagc tgaagagtgg ggatgcttgc attgcagaat atcagcaggc agggagagga
      481 agcagaggga ggaaatggtt ctctcctttt ggagctaacc tgtacctgag tatgttttgg
      541 cgcctgaagc ggggaccagc agcaatcggc ctgggcccgg tcatcggaat tgtcatggca
      601 gaagcgctgc gaaagctggg agcagacaag gtgcgagtca aatggcccaa tgacctgtat
      661 ctgcaggata gaaagctggc aggcatcctg gtggagctgg ccggaataac aggcgatgct
      721 gcacagatcg tcattggcgc cgggattaac gtggctatga ggcgcgtgga ggaaagcgtg
      781 gtcaatcagg gctggatcac actgcaggaa gcagggatta acctggacag gaatactctg
      841 gccgctacgc tgatccgaga gctgcgggca gccctggaac tgttcgagca ggaaggcctg
      901 gctccatatc tgccacggtg ggagaagctg gataacttca tcaatagacc cgtgaagctg
      961 atcattgggg acaaagagat tttcgggatt agccggggga ttgataaaca gggagccctg
     1021 ctgctggaac aggacggagt tatcaaaccc tggatgggcg gagaaatcag tctgcggtct
     1081 gccgaaaaga agcttgcggc cgcactcgag ggaggtggat ctggtggagg atctggagga
     1141 ggttcaatgt gagaccacga gtgattaatg aatcggccaa cgcgcgggga gaggcggttt
     1201 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     1261 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     1321 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1381 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1441 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1501 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1561 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1621 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1681 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1741 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1801 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     1861 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1921 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1981 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2041 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2101 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttattt
     2161 gccgactacc ttggtgatct cgccttttac gtaatggaca aattcttcca actgatctgc
     2221 gcgcgaggcc aagcgatctt cttcttgtcc aagataagcc tgtctagctt caagtatgac
     2281 gggctgatac tgggccggca ggcgctccat tgcccagtcg gcagcgacat ccttcggcgc
     2341 gattttgccg gttactgcgc tgtaccaaat gcgggacaac gtaagcacta catttcgctc
     2401 atcgccagcc cagtcgggcg gcgagttcca tagcgttaag gtttcattta gcgcctcaaa
     2461 tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc gctggaccta ccaaggcaac
     2521 gctatgttct cttgcttttg tcagcaagat agccagatca atgtcgatcg tggctggctc
     2581 gaagatacct gcaagaatgt cattgcgctg ccattctcca aattgcagtt cgcgcttagc
     2641 tggataacgc cacggaatga tgtcgtcgtg cacaacaatg gtgacttcta cagcgcggag
     2701 aatctcgctc tctccagggg aagccgaagt ttccaaaagg tcgttgatca aagctcgccg
     2761 cgttgtttca tcaagcctta cggtcaccgt aaccagcaaa tcaatatcgc tgtgtggctt
     2821 caggccgcca tccactgcgg agccgtacaa atgtacggcc agcaacgtcg gttcgagatg
     2881 gcgctcgatg acgccaacta cctctgatag ttgagtcgat acttcggcga tcaccgcttc
     2941 cctcatgatg tttaactttg ttttagggcg actgccctgc tgcgtaacat cgttgctgct
     3001 ccataacatc aaacatcgac ccacggcgta acgcgcttgc tgcttggatg cccgaggcat
     3061 agactgtacc ccaaaaaaaa acagtcataa caagccatga aaaccgccac tgcgccgtta
     3121 ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg agcgcatact cttccttttt
     3181 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     3241 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     3301 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     3361 tttcgtccac tgaagagcca ctgt
//
