LOCUS       sequence-75093-e        4568 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 4568)
  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 4568)
  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: pICSL70008
FEATURES             Location/Qualifiers
     source          1..4568
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(40..58)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        126..230
                     /gene="bla"
                     /label=AmpR promoter
     CDS             450..1241
                     /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      1412..2000
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1901..1920
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2154..2171
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             3431..4126
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSDMALK
                     LVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHK"
ORIGIN
        1 cgcttgagac cacgaagtgg ctcttcagtg gacgaaaggg cctcgtgata cgcctatttt
       61 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
      121 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
      181 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgcgctcac
      241 gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca
      301 tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag
      361 cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg
      421 gcagtcgccc taaaacaaag ttaaacatca tgagggaagc ggtgatcgcc gaagtatcga
      481 ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg
      541 tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagcgat attgatttgc
      601 tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt
      661 tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg
      721 ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag
      781 aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc
      841 tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg
      901 aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa
      961 cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt
     1021 cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact
     1081 gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt
     1141 atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc
     1201 attacgtaaa aggcgagatc accaaggtag tcggcaaata actgtcagac caagtttact
     1261 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1321 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1381 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1441 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1501 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1561 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1621 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1681 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1741 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1801 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1861 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1921 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1981 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     2041 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     2101 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     2161 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2221 cgattcatta atcactctgt ggtctcagga gttctagaat gtcgcggaac aaattttaaa
     2281 actaaatcct aaatttttct aattttgttg ccaatagtgg atatgtgggc cgtatagaag
     2341 gaatctattg aaggcccaaa cccatactga cgagcccaaa ggttcgtttt gcgttttatg
     2401 tttcggttcg atgccaacgc cacattctga gctaggcaaa aaacaaacgt gtctttgaat
     2461 agactcctct cgttaacaca tgcagcggct gcatggtgac gccattaaca cgtggcctac
     2521 aattgcatga tgtctccatt gacacgtgac ttctcgtctc ctttcttaat atatctaaca
     2581 aacactccta cctcttccaa aatatataca catctttttg atcaatctct cattcaaaat
     2641 ctcattctct ctagtaaaca agaacaaaaa aatggcggat acagctagag gaacccatca
     2701 cgatatcatc ggcagagacc agtacccgat gatgggccga gaccgagacc agtaccagat
     2761 gtccggacga ggatctgact actccaagtc taggcagatt gctaaagctg caactgctgt
     2821 cacagctggt ggttccctcc ttgttctctc cagccttacc cttgttggaa ctgtcatagc
     2881 tttgactgtt gcaacacctc tgctcgttat cttcagccca atccttgtcc cggctctcat
     2941 cacagttgca ctcctcatca ccggttttct ttcctctgga gggtttggca ttgccgctat
     3001 aaccgttttc tcttggattt acaagtaagc acacatttat catcttactt cataattttg
     3061 tgcaatatgt gcatgcatgt gttgagccag tagctttgga tcaatttttt tggtcgaata
     3121 acaaatgtaa caataagaaa ttgcaaattc tagggaacat ttggttaact aaatacgaaa
     3181 tttgacctag ctagcttgaa tgtgtctgtg tatatcatct atataggtaa aatgcttggt
     3241 atgataccta ttgattgtga ataggtacgc aacgggagag cacccacagg gatcagacaa
     3301 gttggacagt gcaaggatga agttgggaag caaagctcag gatctgaaag acagagctca
     3361 gtactacgga cagcaacata ctggtgggga acatgaccgt gaccgtactc gtggtggcca
     3421 gcacactact atgagcgagc tgattaagga gaacatgcac atgaagctgt acatggaggg
     3481 caccgtgaac aaccaccact tcaagtgcac atccgagggc gaaggcaagc cctacgaggg
     3541 cacccagacc atgagaatca aggtggtcga gggcggccct ctccccttcg ccttcgacat
     3601 cctggctacc agcttcatgt acggcagcag aaccttcatc aaccacaccc agggcatccc
     3661 cgacttcttt aagcagtcct tccctgaggg cttcacatgg gagagagtca ccacatacga
     3721 agacgggggc gtgctgaccg ctacccagga caccagcctc caggacggct gcctcatcta
     3781 caacgtcaag atcagagggg tgaacttccc atccaacggc cctgtgatgc agaagaaaac
     3841 actcggctgg gaggccaaca ccgagatgct gtaccccgct gacggcggcc tggaaggcag
     3901 aagcgacatg gccctgaagc tcgtgggcgg gggccacctg atctgcaact tcaagaccac
     3961 atacagatcc aagaaacccg ctaagaacct caagatgccc ggcgtctact atgtggacca
     4021 cagactggaa agaatcaagg aggccgacaa agagacctac gtcgagcagc acgaggtggc
     4081 tgtggccaga tactgcgacc tccctagcaa actggggcac aagtgagctt accccactga
     4141 tgtcatcgtc atagtccaat aactccaatg tcggggagtt agtttatgag gaataaagtg
     4201 tttagaattt gatcaggggg agataataaa agccgagttt gaatcttttt gttataagta
     4261 atgtttatgt gtgtttctat atgttgtcaa atggtaccat gttttttttc ctctcttttt
     4321 gtaacttgca agtgttgtgt tgtactttat ttggcttctt tgtaagttgg taacggtggt
     4381 ctatatatgg aaaaggtctt gttttgttaa acttatgtta gttaactgga ttcgtcttta
     4441 accacaaaaa gttttcaata agctacaaat ttagacacgc aagccgatgc agtcattagt
     4501 acatatattt attgcaagtg attacatggc aacccaaact tcaaaaacag taggttgctc
     4561 catttagt
//
