LOCUS       sequence-499922-        2802 bp    DNA     circular SYN 12-FEB-2026
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2802)
  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 2802)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 12, 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: pICH42222
FEATURES             Location/Qualifiers
     source          1..2802
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..131
                     /gene="bla"
                     /label=AmpR promoter
     CDS             353..1144
                     /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      1315..1903
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1804..1823
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2057..2074
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             2152..2703
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     primer_bind     complement(2743..2761)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
       61 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
      121 aaaggaagag tatgcgctca cgcaactggt ccagaacctt gaccgaacgc agcggtggta
      181 acggcgcagt ggcggttttc atggcttgtt atgactgttt ttttttgggg tacagtctat
      241 gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag
      301 cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca tcatgaggga
      361 agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca
      421 tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa
      481 gccacacagc gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg
      541 gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct
      601 ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc
      661 taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga
      721 gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt
      781 tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt
      841 tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga
      901 gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc
      961 gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt
     1021 catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc
     1081 agatcagttg gaagaatttg tccattacgt aaaaggcgag atcaccaagg tagtcggcaa
     1141 ataactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     1201 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     1261 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     1321 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     1381 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     1441 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     1501 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     1561 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     1621 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     1681 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     1741 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     1801 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     1861 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     1921 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     1981 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     2041 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     2101 accgcctctc cccgcgcgtt ggccgattca ttaatcactc tgtggtctca aatgagccca
     2161 gaacgacgcc cggccgacat ccgccgtgcc accgaggcgg acatgccggc ggtctgcacc
     2221 atcgtcaacc actacatcga gacaagcacg gtcaacttcc gtaccgagcc gcaggaaccg
     2281 caggagtgga cggacgacct cgtccgtctg cgggagcgct atccctggct cgtcgccgag
     2341 gtggacggcg aggtcgccgg catcgcctac gcgggcccct ggaaggcacg caacgcctac
     2401 gactggacgg ccgagtcaac cgtgtacgta tccccccgcc accagcggac gggactgggc
     2461 tccacgctct acacccacct gctgaagtcc ctggaggcac agggcttcaa gagcgtggtc
     2521 gctgtcatcg ggctgcccaa cgacccgagc gtgcgcatgc acgaggcgct cggatatgcc
     2581 ccccgcggca tgctgcgggc ggccggcttc aagcacggga actggcatga cgtgggtttc
     2641 tggcagctgg acttcagcct gccggtaccg ccccgtccgg tcctgcccgt caccgagatt
     2701 tgagctttga gaccacgaag tggctcttca gtggacgaaa gggcctcgtg atacgcctat
     2761 ttttataggt taatgtcatg ataataatgg tttcttagac gt
//
