LOCUS       sequence-495481-        2985 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 2985)
  TITLE     Plant Golden-gate N-Terminal and C-Terminal Tags.
REFERENCE   2  (bases 1 to 2985)
  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: pICSL30053
FEATURES             Location/Qualifiers
     source          1..2985
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(10..12,13..15,16..726)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(55..76)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(316..335)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     663..684
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             727..747
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(826..843)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(997..1585)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1077..1096)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1756..2547)
                     /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(2769..2873)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2941..2959
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ggtctcacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
       61 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
      121 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
      181 tggcccaccc tcgtgaccac cttcagctac ggcgtgcagt gtttcagccg ctaccccgac
      241 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
      301 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
      361 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
      421 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
      481 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
      541 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
      601 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
      661 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctcacggcat ggacgagctg
      721 tacaaggaga atttgtattt tcagtcaatg tgagaccacg agtgattaat gaatcggcca
      781 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
      841 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
      901 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
      961 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1021 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1081 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1141 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     1201 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1261 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     1321 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1381 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     1441 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1501 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1561 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1621 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     1681 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1741 aacttggtct gacagttatt tgccgactac cttggtgatc tcgcctttta cgtaatggac
     1801 aaattcttcc aactgatctg cgcgcgaggc caagcgatct tcttcttgtc caagataagc
     1861 ctgtctagct tcaagtatga cgggctgata ctgggccggc aggcgctcca ttgcccagtc
     1921 ggcagcgaca tccttcggcg cgattttgcc ggttactgcg ctgtaccaaa tgcgggacaa
     1981 cgtaagcact acatttcgct catcgccagc ccagtcgggc ggcgagttcc atagcgttaa
     2041 ggtttcattt agcgcctcaa atagatcctg ttcaggaacc ggatcaaaga gttcctccgc
     2101 cgctggacct accaaggcaa cgctatgttc tcttgctttt gtcagcaaga tagccagatc
     2161 aatgtcgatc gtggctggct cgaagatacc tgcaagaatg tcattgcgct gccattctcc
     2221 aaattgcagt tcgcgcttag ctggataacg ccacggaatg atgtcgtcgt gcacaacaat
     2281 ggtgacttct acagcgcgga gaatctcgct ctctccaggg gaagccgaag tttccaaaag
     2341 gtcgttgatc aaagctcgcc gcgttgtttc atcaagcctt acggtcaccg taaccagcaa
     2401 atcaatatcg ctgtgtggct tcaggccgcc atccactgcg gagccgtaca aatgtacggc
     2461 cagcaacgtc ggttcgagat ggcgctcgat gacgccaact acctctgata gttgagtcga
     2521 tacttcggcg atcaccgctt ccctcatgat gtttaacttt gttttagggc gactgccctg
     2581 ctgcgtaaca tcgttgctgc tccataacat caaacatcga cccacggcgt aacgcgcttg
     2641 ctgcttggat gcccgaggca tagactgtac cccaaaaaaa aacagtcata acaagccatg
     2701 aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt
     2761 gagcgcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     2821 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     2881 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     2941 aataggcgta tcacgaggcc ctttcgtcca ctgaagagcc actgt
//
