LOCUS       sequence-495471-        2964 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 2964)
  TITLE     Plant Golden-gate N-Terminal and C-Terminal Tags.
REFERENCE   2  (bases 1 to 2964)
  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: pICSL30032
FEATURES             Location/Qualifiers
     source          1..2964
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(10..12,13..15,16..732)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYKM"
     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"
     primer_bind     complement(805..822)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(976..1564)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1056..1075)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1735..2526)
                     /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(2748..2852)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2920..2938
                     /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 cctgacctac ggcgtgcagt gcttcagccg 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 ccagtccaag ctgagcaaag accccaacga gaagcgcgat
      661 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctcacggcat ggacgagctg
      721 tacaaaatgt gagaccacga gtgattaatg aatcggccaa cgcgcgggga gaggcggttt
      781 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
      841 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
      901 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      961 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1021 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1081 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1141 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1201 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1261 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1321 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1381 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     1441 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1501 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1561 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     1621 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     1681 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttattt
     1741 gccgactacc ttggtgatct cgccttttac gtaatggaca aattcttcca actgatctgc
     1801 gcgcgaggcc aagcgatctt cttcttgtcc aagataagcc tgtctagctt caagtatgac
     1861 gggctgatac tgggccggca ggcgctccat tgcccagtcg gcagcgacat ccttcggcgc
     1921 gattttgccg gttactgcgc tgtaccaaat gcgggacaac gtaagcacta catttcgctc
     1981 atcgccagcc cagtcgggcg gcgagttcca tagcgttaag gtttcattta gcgcctcaaa
     2041 tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc gctggaccta ccaaggcaac
     2101 gctatgttct cttgcttttg tcagcaagat agccagatca atgtcgatcg tggctggctc
     2161 gaagatacct gcaagaatgt cattgcgctg ccattctcca aattgcagtt cgcgcttagc
     2221 tggataacgc cacggaatga tgtcgtcgtg cacaacaatg gtgacttcta cagcgcggag
     2281 aatctcgctc tctccagggg aagccgaagt ttccaaaagg tcgttgatca aagctcgccg
     2341 cgttgtttca tcaagcctta cggtcaccgt aaccagcaaa tcaatatcgc tgtgtggctt
     2401 caggccgcca tccactgcgg agccgtacaa atgtacggcc agcaacgtcg gttcgagatg
     2461 gcgctcgatg acgccaacta cctctgatag ttgagtcgat acttcggcga tcaccgcttc
     2521 cctcatgatg tttaactttg ttttagggcg actgccctgc tgcgtaacat cgttgctgct
     2581 ccataacatc aaacatcgac ccacggcgta acgcgcttgc tgcttggatg cccgaggcat
     2641 agactgtacc ccaaaaaaaa acagtcataa caagccatga aaaccgccac tgcgccgtta
     2701 ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg agcgcatact cttccttttt
     2761 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     2821 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     2881 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     2941 tttcgtccac tgaagagcca ctgt
//
