LOCUS       sequence-75090-e        4873 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 4873)
  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 4873)
  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: pICSL70002
FEATURES             Location/Qualifiers
     source          1..4873
                     /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             3708..4673
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             3708..4670
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     polyA_signal    4676..4850
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
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 gggtaccgac aaaatttaga acgaacttaa
     2281 ttatgatctc aaatacattg atacatatct catctagatc taggttatca ttatgtaaga
     2341 aagttttgac gaatatggca cgacaaaatg gctagactcg atgtaattgg tatctcaact
     2401 caacattata cttataccaa acattagtta gacaaaattt aaacaactat tttttatgta
     2461 tgcaagagtc agcatatgta taattgattc agaatcgttt tgacgagttc ggatgtagta
     2521 gtagccatta tttaatgtac atactaatcg tgaatagtga atatgatgaa acattgtatc
     2581 ttattgtata aatatccata aacacatcat gaaagacact ttctttcacg gtctgaatta
     2641 attatgatac aattctaata gaaaacgaat taaattacgt tgaattgtat gaaatctaat
     2701 tgaacaagcc aaccacgacg acgactaacg ttgcctggat tgactcggtt taagttaacc
     2761 actaaaaaaa cggagctgtc atgtaacacg cggatcgagc aggtcacagt catgaagcca
     2821 tcaaagcaaa agaactaatc caagggctga gatgattaat tagtttaaaa attagttaac
     2881 acgagggaaa aggctgtctg acagccaggt cacgttatct ttacctgtgg tcgaaatgat
     2941 tcgtgtctgt cgattttaat tatttttttg aaaggccgaa aataaagttg taagagataa
     3001 acccgcctat ataaattcat atattttcct ctccgctttg aattgtctcg ttgtcctcct
     3061 cactttcatc agccgttttg aatctccggc gacttgacag agaagaacaa ggaagaagat
     3121 taagagagaa agtaagagat aatccaggag attcattctc cgttttgaat cttcctcaat
     3181 ctcatcttct tccgctcttt ctttccaagg taataggaac tttctggatc tactttattt
     3241 gctggatctc gatcttgttt tctcaatttc cttgagatct ggaattcgtt taatttggat
     3301 ctgtgaacct ccactaaatc ttttggtttt actagaatcg atctaagttg accgatcagt
     3361 tagctcgatt atagctacca gaatttggct tgaccttgat ggagagatcc atgttcatgt
     3421 tacctgggaa atgatttgta tatgtgaatt gaaatctgaa ctgttgaagt tagattgaat
     3481 ctgaacactg tcaatgttag attgaatctg aacactgttt aaggttagat gaagtttgtg
     3541 tatagattct tcgaaacttt aggatttgta gtgtcgtacg ttgaacagaa agctatttct
     3601 gattcaatca gggtttattt gactgtattg aactcttttt gtgtgtttgc agctcataaa
     3661 aaatggctga ggctgatgat attcaaccaa tcgtgtgtga cctcgagatg aaggataaca
     3721 ccgtgccact gaaattgatt gccctgttag cgaacggtga atttcactct ggcgagcagt
     3781 tgggtgaaac gctgggaatg agccgggcgg ctattaataa acacattcag acactgcgtg
     3841 actggggcgt tgatgtcttt accgttccgg gtaaaggata cagcctgcct gagcctatcc
     3901 agttacttaa tgctaaacag atattgggtc agctggatgg cggtagtgta gccgtgctgc
     3961 cagtgattga ctccacgaat cagtaccttc ttgatcgtat cggagagctt aaatcgggcg
     4021 atgcttgcat tgcagaatac cagcaggctg gccgtggtcg ccggggtcgg aaatggtttt
     4081 cgccttttgg cgcaaactta tatttgtcga tgttctggcg tctggaacaa ggcccggcgg
     4141 cggcgattgg tttaagtctg gttatcggta tcgtgatggc ggaagtatta cgcaagctgg
     4201 gtgcagataa agttcgtgtt aaatggccta atgacctcta tctgcaggat cgcaagctgg
     4261 caggcattct ggtggagctg actggcaaaa ctggcgatgc ggcgcaaata gtcattggag
     4321 ccgggatcaa catggcaatg cgccgtgttg aagagagtgt cgttaatcag gggtggatca
     4381 cgctgcagga agcggggatc aatctcgatc gtaatacgtt ggcggccatg ctaatacgtg
     4441 aattacgtgc tgcgttggaa ctcttcgaac aagaaggatt ggcaccttat ctgtcgcgct
     4501 gggaaaagct ggataatttt attaatcgcc cagtgaaact tatcattggt gataaagaaa
     4561 tatttggcat ttcacgcgga atagacaaac agggggcttt attacttgag caggatggaa
     4621 taataaaacc ctggatgggc ggtgaaatat ccctgcgtag tgcagaaaaa taagctttct
     4681 ccataataat gtgtgagtag ttcccagata agggaattag ggttcctata gggtttcgct
     4741 catgtgttga gcatataaga aacccttagt atgtatttgt atttgtaaaa tacttctatc
     4801 aataaaattt ctaattccta aaaccaaaat ccagtactaa aatccagatc cccgtcgacc
     4861 gatatccgaa ttc
//
