LOCUS       sequence-495513-        3280 bp    DNA     circular SYN 17-DEC-2025
DEFINITION  Level 0 Golden Gate part C-Terminal Tag with overhangs TTCG - GCTT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3280)
  TITLE     Plant Golden-gate N-Terminal and C-Terminal Tags.
REFERENCE   2  (bases 1 to 3280)
  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: pICSL50035
FEATURES             Location/Qualifiers
     source          1..3280
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             12..44
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             60..1019
                     /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(1080..1098)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1166..1270
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1492..2283
                     /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"
     rep_origin      2454..3042
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2943..2962
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3196..3213
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggtctcattc gatggctagc atgactggtg gacagcaaat gggtcgcgga tccgaattca
       61 tgaaagacaa tactgtgcct ctgaagctga tcgctctcct ggctaatggc gagttccata
      121 gtggcgaaca gctgggagaa accctgggca tgtccagggc cgctatcaac aagcacattc
      181 agactctgcg cgactggggc gtggacgtgt tcaccgtgcc cggaaagggc tactctctgc
      241 ccgagcctat cccgctgctg aacgctaaac agattttggg acagctggac ggcgggagcg
      301 tggcagtcct gcctgtggtc gactccacca atcagtacct gctggatcga atcggcgagc
      361 tgaagagtgg ggatgcttgc attgcagaat atcagcaggc agggagagga agcagaggga
      421 ggaaatggtt ctctcctttt ggagctaacc tgtacctgag tatgttttgg cgcctgaagc
      481 ggggaccagc agcaatcggc ctgggcccgg tcatcggaat tgtcatggca gaagcgctgc
      541 gaaagctggg agcagacaag gtgcgagtca aatggcccaa tgacctgtat ctgcaggata
      601 gaaagctggc aggcatcctg gtggagctgg ccggaataac aggcgatgct gcacagatcg
      661 tcattggcgc cgggattaac gtggctatga ggcgcgtgga ggaaagcgtg gtcaatcagg
      721 gctggatcac actgcaggaa gcagggatta acctggacag gaatactctg gccgctacgc
      781 tgatccgaga gctgcgggca gccctggaac tgttcgagca ggaaggcctg gctccatatc
      841 tgccacggtg ggagaagctg gataacttca tcaatagacc cgtgaagctg atcattgggg
      901 acaaagagat tttcgggatt agccggggga ttgataaaca gggagccctg ctgctggaac
      961 aggacggagt tatcaaaccc tggatgggcg gagaaatcag tctgcggtct gccgaaaaga
     1021 agcttgcggc cgcactcgag tgagctttga gaccacgtgg ctcttcagtg gacgaaaggg
     1081 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     1141 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     1201 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     1261 aaggaagagt atgcgctcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa
     1321 cggcgcagtg gcggttttca tggcttgtta tgactgtttt tttttggggt acagtctatg
     1381 cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc
     1441 agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaaacat catgagggaa
     1501 gcggtgatcg ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat
     1561 ctcgaaccga cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag
     1621 ccacacagcg atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg
     1681 cgagctttga tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc
     1741 cgcgctgtag aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct
     1801 aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag
     1861 ccagccacga tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt
     1921 gccttggtag gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt
     1981 gaggcgctaa atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag
     2041 cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg
     2101 ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc
     2161 atacttgaag ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca
     2221 gatcagttgg aagaatttgt ccattacgta aaaggcgaga tcaccaaggt agtcggcaaa
     2281 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     2341 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     2401 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2461 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2521 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2581 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     2641 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2701 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2761 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2821 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     2881 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     2941 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3001 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3061 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3121 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3181 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     3241 ccgcctctcc ccgcgcgttg gccgattcat taatcactgt
//
