LOCUS       sequence-75071-e        3563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3563)
  AUTHORS   Engler C, Youles M, Gruetzner R, Ehnert TM, Werner S, Jones JD, 
            Patron NJ, Marillonnet S
  TITLE     A golden gate modular cloning toolbox for plants.
  JOURNAL   ACS Synth Biol. 2014 Nov 21;3(11):839-43. doi: 10.1021/sb4001504. 
            Epub 2014 Feb 20.
  PUBMED    24933124
REFERENCE   2  (bases 1 to 3563)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1021/sb4001504";
            journalName: "ACS Synth Biol"; date: "2014-11-21- 21"; volume: "3"; 
            issue: "11"; pages: "839-43"
COMMENT     Sequence Label: pICSL50008
FEATURES             Location/Qualifiers
     source          1..3563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(63..651)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(143..162)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(744..1535)
                     /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"
     primer_bind     1966..1982
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1966..1982
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1987..2006
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1987..2005
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2024..2123
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(2152..2154,2155..2868)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2194..2215)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2455..2474)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2802..2823
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2896..2995)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3011..3028)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3011..3027)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3020..3042)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      3091..3177
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3269..3296
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3455..3472)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
       61 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      121 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      181 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      241 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      301 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      361 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      421 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      481 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
      541 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      601 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      661 ttgatctttt ctacggggtc tgacgctcag tggaacgacg cgtaactcac gttaagggat
      721 tttggtcatg ggtggctcga gggttatttg ccgactacct tggtgatctc gcctttcacg
      781 tagtggacaa attcttccaa ctgatctgcg cgcgaggcca agcgatcttc ttcttgtcca
      841 agataagcct gtctagcttc aagtatgacg ggctgatact gggccggcag gcgctccatt
      901 gcccagtcgg cagcgacatc cttcggcgcg attttgccgg ttactgcgct gtaccaaatg
      961 cgggacaacg taagcactac atttcgctca tcgccagccc agtcgggcgg cgagttccat
     1021 agcgttaagg tttcatttag cgcctcaaat agatcctgtt caggaaccgg atcaaagagt
     1081 tcctccgccg ctggacctac caaggcaacg ctatgttctc ttgcttttgt cagcaagata
     1141 gccagatcaa tgtcgatcgt ggctggctcg aagatacctg caagaatgtc attgcgctgc
     1201 cattctccaa attgcagttc gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc
     1261 acaacaatgg tgacttctac agcgcggaga atctcgctct ctccagggga agccgaagtt
     1321 tccaaaaggt cgttgatcaa agctcgccgc gttgtttcat caagccttac ggtcaccgta
     1381 accagcaaat caatatcact gtgtggcttc aggccgccat ccactgcgga gccgtacaaa
     1441 tgtacggcca gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac ctctgatagt
     1501 tgagtcgata cttcggcgat caccgcttcc ctcatgatgt ttaactttgt tttagggcga
     1561 ctgccctgct gcgtaacatc gttgctgctc cataacatca aacatcgacc cacggcgtaa
     1621 cgcgcttgct gcttggatgc ccgaggcata gactgtaccc caaaaaaaca gtcataacaa
     1681 gccatgaaaa ccgccaccgc gccgttacca ccgctgcgtt cggtcaaggt tctggaccag
     1741 ttgcgtgagc gcatacgcta cttgcattac agcttacgaa ccgaacaggc ttatgtccac
     1801 tgggttcgtg ccttcatccg tttccacggt gtgcgtcacc cggcaacctt gggtagcagc
     1861 gaagtcgagg catttctgtc ctggctggtc tagaggaggc atgatgatat atttttatct
     1921 tgtgcaatgt aacatcagag attttgagac acgggccaga gctgccagga aacagctatg
     1981 accatgtaat acgactcact ataggggata tcagctggat ggcaaataat gatttttatt
     2041 ttgactgata gtgacctgtt cgttgcaaca aattgatgag caattatttt ttataatgcc
     2101 aactttgtac aagaaagctg ggtcgaattc gcccttcacc ggtctctttc ggtgagcaag
     2161 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     2221 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     2281 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     2341 ttcagctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     2401 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     2461 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     2521 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     2581 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     2641 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     2701 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     2761 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     2821 gtgaccgccg ccgggatcac tcacggcatg gacgagctgt acaagtaagc tttgagacct
     2881 aaagggcgaa ttcggagcct gcttttttgt acaaagttgg cattataaaa aagcattgct
     2941 catcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttggggcc
     3001 cgagcttaag actggccgtc gttttacaac gtcgtgactg ggaaaacatc catgctagcg
     3061 ttaacgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca gtcggaagac
     3121 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
     3181 ccgggagcgg atttgaacgt tgtgaagcaa cggcccggag ggtggcgggc aggacgcccg
     3241 ccataaactg ccaggcatca aactaagcag aaggccatcc tgacggatgg cctttttgcg
     3301 tttctacaaa ctcttcctgg ctagcggtac gcgtattaat tgcgttgcgc tcactgcccg
     3361 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3421 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3481 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3541 aatcagggga taacgcagga aag
//
