LOCUS       sequence-495577-        3361 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 3361)
  TITLE     Plant Golden-gate N-Terminal and C-Terminal Tags.
REFERENCE   2  (bases 1 to 3361)
  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: pICSL50040
FEATURES             Location/Qualifiers
     source          1..3361
                     /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"
     CDS             1098..1121
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1161..1179)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1247..1351
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1573..2364
                     /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      2535..3123
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3024..3043
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3277..3294
                     /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 gattataagg accatgacgg agactataag gaccatgacc
     1081 tcgacgctgc agcagctgat tataaggacg atgacgataa gtgagctttg agaccacgtg
     1141 gctcttcagt ggacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     1201 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     1261 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     1321 aatgcttcaa taatattgaa aaaggaagag tatgcgctca cgcaactggt ccagaacctt
     1381 gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt atgactgttt
     1441 ttttttgggg tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt
     1501 cgatgtttga tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca
     1561 aagttaaaca tcatgaggga agcggtgatc gccgaagtat cgactcaact atcagaggta
     1621 gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc
     1681 gcagtggatg gcggcctgaa gccacacagc gatattgatt tgctggttac ggtgaccgta
     1741 aggcttgatg aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc
     1801 cctggagaga gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc
     1861 attccgtggc gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac
     1921 attcttgcag gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca
     1981 aaagcaagag aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg
     2041 gttcctgaac aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg
     2101 cccgactggg ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc
     2161 gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg
     2221 ccggcccagt atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa
     2281 gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg tccattacgt aaaaggcgag
     2341 atcaccaagg tagtcggcaa ataactgtca gaccaagttt actcatatat actttagatt
     2401 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     2461 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     2521 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2581 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2641 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     2701 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2761 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2821 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     2881 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     2941 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3001 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3061 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3121 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3181 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     3241 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     3301 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatcactg
     3361 t
//
