LOCUS       sequence-75085-e        3407 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 3407)
  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 3407)
  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: pICH42222
FEATURES             Location/Qualifiers
     source          1..3407
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..131
                     /gene="bla"
                     /label=AmpR promoter
     CDS             351..1142
                     /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      1313..1901
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1802..1821
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2055..2072
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2189..2210
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2225..2230,2231..2248,2249..2255)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2263..2279
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2268..2290
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2287..2303
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2287..2303
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2362..2913
                     /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(2944..2961)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2944..2960)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2953..2975)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3169..3188)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3288..3310
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3348..3366)
                     /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 ttttggggta cagtctatgc
      241 ctcgggcatc caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca
      301 gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa gttaaacatc atgagggaag
      361 cggtgatcgc cgaagtatcg actcaactat cagaggtagt tggcgtcatc gagcgccatc
      421 tcgaaccgac gttgctggcc gtacatttgt acggctccgc agtggatggc ggcctgaagc
      481 cacacagcga tattgatttg ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc
      541 gagctttgat caacgacctt ttggaaactt cggcttcccc tggagagagc gagattctcc
      601 gcgctgtaga agtcaccatt gttgtgcacg acgacatcat tccgtggcgt tatccagcta
      661 agcgcgaact gcaatttgga gaatggcagc gcaatgacat tcttgcaggt atcttcgagc
      721 cagccacgat cgacattgat ctggctatct tgctgacaaa agcaagagaa catagcgttg
      781 ccttggtagg tccagcggcg gaggaactct ttgatccggt tcctgaacag gatctatttg
      841 aggcgctaaa tgaaacctta acgctatgga actcgccgcc cgactgggct ggcgatgagc
      901 gaaatgtagt gcttacgttg tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc
      961 cgaaggatgt cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca
     1021 tacttgaagc tagacaggct tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag
     1081 atcagttgga agaatttgtc cattacgtaa aaggcgagat caccaaggta gtcggcaaat
     1141 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     1201 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     1261 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     1321 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     1381 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     1441 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     1501 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     1561 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     1621 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     1681 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     1741 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     1801 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     1861 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     1921 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     1981 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     2041 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     2101 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     2161 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     2221 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     2281 ttcacacagg aaacagctat gaccatgatt acgccaagct tgcatgcctg caggtcgact
     2341 ctagaggatc cccggtctca aatgagccca gaacgacgcc cggccgacat ccgccgtgcc
     2401 accgaggcgg acatgccggc ggtctgcacc atcgtcaacc actacatcga gacaagcacg
     2461 gtcaacttcc gtaccgagcc gcaggaaccg caggagtgga cggacgacct cgtccgtctg
     2521 cgggagcgct atccctggct cgtcgccgag gtggacggcg aggtcgccgg catcgcctac
     2581 gcgggcccct ggaaggcacg caacgcctac gactggacgg ccgagtcaac cgtgtacgta
     2641 tccccccgcc accagcggac gggactgggc tccacgctct acacccacct gctgaagtcc
     2701 ctggaggcac agggcttcaa gagcgtggtc gctgtcatcg ggctgcccaa cgacccgagc
     2761 gtgcgcatgc acgaggcgct cggatatgcc ccccgcggca tgctgcgggc ggccggcttc
     2821 aagcacggga actggcatga cgtgggtttc tggcagctgg acttcagcct gccggtaccg
     2881 ccccgtccgg tcctgcccgt caccgagatt tgagctttga gaccgggtac cgagctcgaa
     2941 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     3001 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     3061 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct
     3121 ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc
     3181 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     3241 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accatctccg ggagctgcat
     3301 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagt cgaaagggcc tcgtgatacg
     3361 cctattttta taggttaatg tcatgataat aatggtttct tagacgt
//
