LOCUS       sequence-495527-        3157 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 3157)
  TITLE     Plant Golden-gate N-Terminal and C-Terminal Tags.
REFERENCE   2  (bases 1 to 3157)
  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: pICSL50062
FEATURES             Location/Qualifiers
     source          1..3157
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(12..14,15..725)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(54..75)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(315..334)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     662..683
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             726..815
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGLYPYDVPDYA"
     CDS             828..854
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             858..884
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             891..917
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(957..975)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1043..1147
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1369..2160
                     /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      2331..2919
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2820..2839
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3073..3090
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggtctcattc ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
       61 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      121 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
      181 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
      241 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      301 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      361 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
      421 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
      481 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
      541 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
      601 acaaccacta cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
      661 acatggtcct gctggagttc gtgaccgccg ccgggatcac tcacggcatg gacgagctgt
      721 acaagtaccc atacgatgtt cctgactatg cgggctatcc ctatgacgtc ccggactatg
      781 caggattgta tccatatgac gttccagatt acgccactag agctgcttac ccatacgatg
      841 ttcctgacta tgcgggctat ccctatgacg tcccggacta tgcaggattg tacccgtacg
      901 atgtgcccga ctacgcttga gctttgagac cacgtggctc ttcagtggac gaaagggcct
      961 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     1021 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     1081 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     1141 gaagagtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg gtggtaacgg
     1201 cgcagtggcg gttttcatgg cttgttatga ctgttttttt ttggggtaca gtctatgcct
     1261 cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc
     1321 aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taaacatcat gagggaagcg
     1381 gtgatcgccg aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc
     1441 gaaccgacgt tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca
     1501 cacagcgata ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga
     1561 gctttgatca acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc
     1621 gctgtagaag tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag
     1681 cgcgaactgc aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca
     1741 gccacgatcg acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc
     1801 ttggtaggtc cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag
     1861 gcgctaaatg aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga
     1921 aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg
     1981 aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata
     2041 cttgaagcta gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat
     2101 cagttggaag aatttgtcca ttacgtaaaa ggcgagatca ccaaggtagt cggcaaataa
     2161 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     2221 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     2281 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2341 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2401 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2461 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     2521 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2581 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2641 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2701 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     2761 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     2821 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     2881 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     2941 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3001 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3061 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     3121 cctctccccg cgcgttggcc gattcattaa tcactgt
//
