LOCUS       sequence-495578-        2464 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 2464)
  TITLE     Plant Golden-gate N-Terminal and C-Terminal Tags.
REFERENCE   2  (bases 1 to 2464)
  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: pICSL50041
FEATURES             Location/Qualifiers
     source          1..2464
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             12..77
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             78..101
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             102..131
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             141..164
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             201..224
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(264..282)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        350..454
                     /gene="bla"
                     /label=AmpR promoter
     CDS             676..1467
                     /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      1638..2226
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2127..2146
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2380..2397
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggtctcattc ggactacaaa gaccatgacg gtgattataa agatcatgat atcgattaca
       61 aggatgacga tgacaaactg gaagttctgt tccaggggcc acatcaccac caccatcatc
      121 atcaccatca cggctccggc tggagccacc cgcagttcga aaaaggcgga ggttccggcg
      181 gaggttctgg tggaggctcc tggagtcatc ctcaatttga gaagtaggct ttgagaccac
      241 gtggctcttc agtggacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
      301 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
      361 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
      421 ataaatgctt caataatatt gaaaaaggaa gagtatgcgc tcacgcaact ggtccagaac
      481 cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt gttatgactg
      541 tttttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg
      601 ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa
      661 acaaagttaa acatcatgag ggaagcggtg atcgccgaag tatcgactca actatcagag
      721 gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc
      781 tccgcagtgg atggcggcct gaagccacac agcgatattg atttgctggt tacggtgacc
      841 gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct
      901 tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac
      961 atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat
     1021 gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg
     1081 acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat
     1141 ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg
     1201 ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac
     1261 agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc
     1321 ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa
     1381 gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccatta cgtaaaaggc
     1441 gagatcacca aggtagtcgg caaataactg tcagaccaag tttactcata tatactttag
     1501 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     1561 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1621 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1681 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     1741 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     1801 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     1861 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     1921 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     1981 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2041 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2101 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2161 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2221 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2281 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     2341 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     2401 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatca
     2461 ctgt
//
