LOCUS       sequence-75091-e        4118 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 4118)
  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 4118)
  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: pICSL70004
FEATURES             Location/Qualifiers
     source          1..4118
                     /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"
     promoter        2326..2505
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    2536..2590
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             2592..3386
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MVEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2646..2665)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3256..3275
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
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 gcggagaatt aagggagtca cgttatgacc
     2281 cccgccgatg acgcgggaca agccgtttta cgtttggaac tgacagaacc gcaacgttga
     2341 aggagccact gagccgcggg tttctggagt ttaatgagct aagcacatac gtcagaaacc
     2401 attattgcgc gttcaaaagt cgcctaaggt cactatcagc tagcaaatat ttcttgtcaa
     2461 aaatgctcca ctgacgttcc ataaattccc ctcggtatcc aattagagtc tcatattcac
     2521 tctcctattt ttacaacaat taccaacaac aacaaacaac aaacaacatt acaattacat
     2581 ttacaattac catggttgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     2641 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     2701 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     2761 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     2821 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     2881 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     2941 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     3001 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     3061 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     3121 gcatgcccga cggcgaggat ctcgtcgtga ctcatggcga tgcctgcttg ccgaatatca
     3181 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     3241 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     3301 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     3361 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgctagag tcctgcttta
     3421 atgagatatg cgagacgcct atgatcgcat gatatttgct ttcaattctg ttgtgcacgt
     3481 tgtaaaaaac ctgagcatgt gtagctcaga tccttaccgc cggtttcggt tcattctaat
     3541 gaatatatca cccgttacta tcgtattttt atgaataata ttctccgttc aatttactga
     3601 ttgtacccta ctacttatat gtacaatatt aaaatgaaaa caatatattg tgctgaatag
     3661 gtttatagcg acatctatga tagagcgcca caataacaaa caattgcgtt ttattattac
     3721 aaatccaatt ttaaaaaaag cggcagaacc ggtcaaacct aaaagactga ttacataaat
     3781 cttattcaaa tttcaaaagt gccccagggg ctagtatcta cgacacaccg agcggcgaac
     3841 taataacgct cactgaaggg aactccggtt ccccgccggc gcgcatgggt gagattcctt
     3901 gaagttgagt attggccgtc cgctctaccg aaagttacgg gcaccattca acccggtcca
     3961 gcacggcggc cgggtaaccg acttgctgcc ccgagaatta tgcagcattt ttttggtgta
     4021 tgtgggcccc aaatgaagtg caggtcaaac cttgacagtg acgacaaatc gttgggcggg
     4081 tccagggcga attttgcgac aacatgtcga ggctcagc
//
