LOCUS       sequence-75061-e        2971 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 2971)
  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 2971)
  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: pICSL30003
FEATURES             Location/Qualifiers
     source          1..2971
                     /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             1397..2104
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1542..1560)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(2201..2218)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2372..2960)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2452..2471)
                     /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 cgaggaggat aacatggcca tcatcaagga
     1441 gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga
     1501 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
     1561 gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa
     1621 ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg
     1681 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga
     1741 ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc
     1801 ctccgacggc cccgtaatgc agaagaaaac catgggctgg gaggcctcct ccgagcggat
     1861 gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg
     1921 cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc
     1981 cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat
     2041 cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta
     2101 caagaacgcg gctattagat caatgtgaga ccacgagtga ttaatgaatc ggccaacgcg
     2161 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     2221 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2281 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2341 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2401 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2461 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2521 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2581 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2641 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2701 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2761 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     2821 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2881 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2941 gcagaaaaaa aggatctcaa gaagatcctt t
//
