LOCUS       sequence-495565-        3363 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 3363)
  TITLE     Plant Golden-gate N-Terminal and C-Terminal Tags.
REFERENCE   2  (bases 1 to 3363)
  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: pICSL30069
FEATURES             Location/Qualifiers
     source          1..3363
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(13..18,19..42,43..78,79..102)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     CDS             109..1068
                     /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(1204..1221)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1375..1963)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1455..1474)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2134..2925)
                     /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(3147..3251)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3319..3337
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ggtctcacca tgtcagcttg gtcccaccct caattcgaga aaggaggcgg ttcaggtggc
       61 ggatcggggg gttctgcatg gtcgcatcct cagtttgaaa agtcaggaat gaaagacaat
      121 actgtgcctc tgaagctgat cgctctcctg gctaatggcg agttccatag tggcgaacag
      181 ctgggagaaa ccctgggcat gtccagggcc gctatcaaca agcacattca gactctgcgc
      241 gactggggcg tggacgtgtt caccgtgccc ggaaagggct actctctgcc cgagcctatc
      301 ccgctgctga acgctaaaca gattttggga cagctggacg gcgggagcgt ggcagtcctg
      361 cctgtggtcg actccaccaa tcagtacctg ctggatcgaa tcggcgagct gaagagtggg
      421 gatgcttgca ttgcagaata tcagcaggca gggagaggaa gcagagggag gaaatggttc
      481 tctccttttg gagctaacct gtacctgagt atgttttggc gcctgaagcg gggaccagca
      541 gcaatcggcc tgggcccggt catcggaatt gtcatggcag aagcgctgcg aaagctggga
      601 gcagacaagg tgcgagtcaa atggcccaat gacctgtatc tgcaggatag aaagctggca
      661 ggcatcctgg tggagctggc cggaataaca ggcgatgctg cacagatcgt cattggcgcc
      721 gggattaacg tggctatgag gcgcgtggag gaaagcgtgg tcaatcaggg ctggatcaca
      781 ctgcaggaag cagggattaa cctggacagg aatactctgg ccgctacgct gatccgagag
      841 ctgcgggcag ccctggaact gttcgagcag gaaggcctgg ctccatatct gccacggtgg
      901 gagaagctgg ataacttcat caatagaccc gtgaagctga tcattgggga caaagagatt
      961 ttcgggatta gccgggggat tgataaacag ggagccctgc tgctggaaca ggacggagtt
     1021 atcaaaccct ggatgggcgg agaaatcagt ctgcggtctg ccgaaaagaa gcttgcggcc
     1081 gcactcgagg gaggtggatc tggtggagga tctggaggag gttcaatgtg agaccacgag
     1141 tgattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     1201 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     1261 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     1321 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1381 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1441 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1501 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1561 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1621 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     1681 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     1741 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     1801 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     1861 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     1921 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     1981 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2041 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2101 ctaaagtata tatgagtaaa cttggtctga cagttatttg ccgactacct tggtgatctc
     2161 gccttttacg taatggacaa attcttccaa ctgatctgcg cgcgaggcca agcgatcttc
     2221 ttcttgtcca agataagcct gtctagcttc aagtatgacg ggctgatact gggccggcag
     2281 gcgctccatt gcccagtcgg cagcgacatc cttcggcgcg attttgccgg ttactgcgct
     2341 gtaccaaatg cgggacaacg taagcactac atttcgctca tcgccagccc agtcgggcgg
     2401 cgagttccat agcgttaagg tttcatttag cgcctcaaat agatcctgtt caggaaccgg
     2461 atcaaagagt tcctccgccg ctggacctac caaggcaacg ctatgttctc ttgcttttgt
     2521 cagcaagata gccagatcaa tgtcgatcgt ggctggctcg aagatacctg caagaatgtc
     2581 attgcgctgc cattctccaa attgcagttc gcgcttagct ggataacgcc acggaatgat
     2641 gtcgtcgtgc acaacaatgg tgacttctac agcgcggaga atctcgctct ctccagggga
     2701 agccgaagtt tccaaaaggt cgttgatcaa agctcgccgc gttgtttcat caagccttac
     2761 ggtcaccgta accagcaaat caatatcgct gtgtggcttc aggccgccat ccactgcgga
     2821 gccgtacaaa tgtacggcca gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac
     2881 ctctgatagt tgagtcgata cttcggcgat caccgcttcc ctcatgatgt ttaactttgt
     2941 tttagggcga ctgccctgct gcgtaacatc gttgctgctc cataacatca aacatcgacc
     3001 cacggcgtaa cgcgcttgct gcttggatgc ccgaggcata gactgtaccc caaaaaaaaa
     3061 cagtcataac aagccatgaa aaccgccact gcgccgttac caccgctgcg ttcggtcaag
     3121 gttctggacc agttgcgtga gcgcatactc ttcctttttc aatattattg aagcatttat
     3181 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     3241 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     3301 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtccact gaagagccac
     3361 tgt
//
