LOCUS       sequence-497539-        4120 bp    DNA     circular SYN 15-JAN-2026
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4120)
  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 4120)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 15, 2026 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..4120
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    180..234
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             236..1030
                     /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(290..309)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     900..919
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      1057..1764
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     primer_bind     complement(1802..1820)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1888..1992
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2214..3005
                     /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      3176..3764
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3665..3684
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3918..3935
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(4090..4120,1..149)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
ORIGIN
        1 cgggtttctg gagtttaatg agctaagcac atacgtcaga aaccattatt gcgcgttcaa
       61 aagtcgccta aggtcactat cagctagcaa atatttcttg tcaaaaatgc tccactgacg
      121 ttccataaat tcccctcggt atccaattag agtctcatat tcactctcct atttttacaa
      181 caattaccaa caacaacaaa caacaaacaa cattacaatt acatttacaa ttaccatggt
      241 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
      301 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
      361 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
      421 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
      481 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
      541 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
      601 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
      661 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
      721 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
      781 ggatctcgtc gtgactcatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
      841 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
      901 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
      961 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     1021 gttcttctga gcgggactct ggggttcgct agagtcctgc tttaatgaga tatgcgagac
     1081 gcctatgatc gcatgatatt tgctttcaat tctgttgtgc acgttgtaaa aaacctgagc
     1141 atgtgtagct cagatcctta ccgccggttt cggttcattc taatgaatat atcacccgtt
     1201 actatcgtat ttttatgaat aatattctcc gttcaattta ctgattgtac cctactactt
     1261 atatgtacaa tattaaaatg aaaacaatat attgtgctga ataggtttat agcgacatct
     1321 atgatagagc gccacaataa caaacaattg cgttttatta ttacaaatcc aattttaaaa
     1381 aaagcggcag aaccggtcaa acctaaaaga ctgattacat aaatcttatt caaatttcaa
     1441 aagtgcccca ggggctagta tctacgacac accgagcggc gaactaataa cgctcactga
     1501 agggaactcc ggttccccgc cggcgcgcat gggtgagatt ccttgaagtt gagtattggc
     1561 cgtccgctct accgaaagtt acgggcacca ttcaacccgg tccagcacgg cggccgggta
     1621 accgacttgc tgccccgaga attatgcagc atttttttgg tgtatgtggg ccccaaatga
     1681 agtgcaggtc aaaccttgac agtgacgaca aatcgttggg cgggtccagg gcgaattttg
     1741 cgacaacatg tcgaggctca gccgcttgag accacgaagt ggctcttcag tggacgaaag
     1801 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     1861 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     1921 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     1981 aaaaggaaga gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt
     2041 aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttttggg gtacagtcta
     2101 tgcctcgggc atccaagcag caagcgcgtt acgccgtggg tcgatgtttg atgttatgga
     2161 gcagcaacga tgttacgcag cagggcagtc gccctaaaac aaagttaaac atcatgaggg
     2221 aagcggtgat cgccgaagta tcgactcaac tatcagaggt agttggcgtc atcgagcgcc
     2281 atctcgaacc gacgttgctg gccgtacatt tgtacggctc cgcagtggat ggcggcctga
     2341 agccacacag cgatattgat ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc
     2401 ggcgagcttt gatcaacgac cttttggaaa cttcggcttc ccctggagag agcgagattc
     2461 tccgcgctgt agaagtcacc attgttgtgc acgacgacat cattccgtgg cgttatccag
     2521 ctaagcgcga actgcaattt ggagaatggc agcgcaatga cattcttgca ggtatcttcg
     2581 agccagccac gatcgacatt gatctggcta tcttgctgac aaaagcaaga gaacatagcg
     2641 ttgccttggt aggtccagcg gcggaggaac tctttgatcc ggttcctgaa caggatctat
     2701 ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc gcccgactgg gctggcgatg
     2761 agcgaaatgt agtgcttacg ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg
     2821 cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct gccggcccag tatcagcccg
     2881 tcatacttga agctagacag gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg
     2941 cagatcagtt ggaagaattt gtccattacg taaaaggcga gatcaccaag gtagtcggca
     3001 aataactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     3061 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3121 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3181 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3241 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3301 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     3361 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3421 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3481 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3541 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3601 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3661 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3721 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3781 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3841 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     3901 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     3961 aaccgcctct ccccgcgcgt tggccgattc attaatcact ctgtggtctc aggagcggag
     4021 aattaaggga gtcacgttat gacccccgcc gatgacgcgg gacaagccgt tttacgtttg
     4081 gaactgacag aaccgcaacg ttgaaggagc cactgagccg
//
