LOCUS       sequence-75060-e        2980 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 2980)
  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 2980)
  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: pICSL30006
FEATURES             Location/Qualifiers
     source          1..2980
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(160..951)
                     /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(1171..1275)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1343..1361
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             join(1397..1399,1400..1402,1403..2113)
                     /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(1442..1463)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1703..1722)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2050..2071
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2210..2227)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2381..2969)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2461..2480)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
       61 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
      121 atcaatctaa agtatatatg agtaaacttg gtctgacagt tatttgccga ctaccttggt
      181 gatctcgcct tttacgtaat ggacaaattc ttccaactga tctgcgcgcg aggccaagcg
      241 atcttcttct tgtccaagat aagcctgtct agcttcaagt atgacgggct gatactgggc
      301 cggcaggcgc tccattgccc agtcggcagc gacatccttc ggcgcgattt tgccggttac
      361 tgcgctgtac caaatgcggg acaacgtaag cactacattt cgctcatcgc cagcccagtc
      421 gggcggcgag ttccatagcg ttaaggtttc atttagcgcc tcaaatagat cctgttcagg
      481 aaccggatca aagagttcct ccgccgctgg acctaccaag gcaacgctat gttctcttgc
      541 ttttgtcagc aagatagcca gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag
      601 aatgtcattg cgctgccatt ctccaaattg cagttcgcgc ttagctggat aacgccacgg
      661 aatgatgtcg tcgtgcacaa caatggtgac ttctacagcg cggagaatct cgctctctcc
      721 aggggaagcc gaagtttcca aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag
      781 ccttacggtc accgtaacca gcaaatcaat atcgctgtgt ggcttcaggc cgccatccac
      841 tgcggagccg tacaaatgta cggccagcaa cgtcggttcg agatggcgct cgatgacgcc
      901 aactacctct gatagttgag tcgatacttc ggcgatcacc gcttccctca tgatgtttaa
      961 ctttgtttta gggcgactgc cctgctgcgt aacatcgttg ctgctccata acatcaaaca
     1021 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtaccccaaa
     1081 aaaacagtca taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt
     1141 caaggttctg gaccagttgc gtgagcgcat actcttcctt tttcaatatt attgaagcat
     1201 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     1261 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     1321 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc cactgaagag
     1381 ccactgtggt ctcaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     1441 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     1501 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     1561 cgtgccctgg cccaccctcg tgaccacctt cagctacggc gtgcagtgct tcagccgcta
     1621 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     1681 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     1741 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     1801 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     1861 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     1921 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     1981 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     2041 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc acggcatgga
     2101 cgagctgtac aaaaacgcgg ctattagatc aatgtgagac cacgagtgat taatgaatcg
     2161 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     2221 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     2281 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     2341 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     2401 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     2461 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2521 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     2581 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2641 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2701 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     2761 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     2821 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     2881 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     2941 agattacgcg cagaaaaaaa ggatctcaag aagatccttt
//
