LOCUS       sequence-194302-        4875 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 4875)
  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 4875)
  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..4875
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             159..950
                     /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      1121..1709
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1610..1629
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1863..1880
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             3417..4382
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             3417..4379
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     polyA_signal    4385..4559
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(4622..4640)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4708..4812
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gttttcatgg cttgttatga ctgttttttt ttggggtaca gtctatgcct cgggcatcca
       61 agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta
      121 cgcagcaggg cagtcgccct aaaacaaagt taaacatcat gagggaagcg gtgatcgccg
      181 aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt
      241 tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagcgata
      301 ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca
      361 acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag
      421 tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc
      481 aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg
      541 acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc
      601 cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg
      661 aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc
      721 ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg
      781 ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta
      841 gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag
      901 aatttgtcca ttacgtaaaa ggcgagatca ccaaggtagt cggcaaataa ctgtcagacc
      961 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     1021 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     1081 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     1141 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     1201 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     1261 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     1321 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     1381 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     1441 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     1501 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     1561 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     1621 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     1681 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     1741 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     1801 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     1861 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     1921 cgcgttggcc gattcattaa tcactctgtg gtctcaggag ggtaccgaca aaatttagaa
     1981 cgaacttaat tatgatctca aatacattga tacatatctc atctagatct aggttatcat
     2041 tatgtaagaa agttttgacg aatatggcac gacaaaatgg ctagactcga tgtaattggt
     2101 atctcaactc aacattatac ttataccaaa cattagttag acaaaattta aacaactatt
     2161 ttttatgtat gcaagagtca gcatatgtat aattgattca gaatcgtttt gacgagttcg
     2221 gatgtagtag tagccattat ttaatgtaca tactaatcgt gaatagtgaa tatgatgaaa
     2281 cattgtatct tattgtataa atatccataa acacatcatg aaagacactt tctttcacgg
     2341 tctgaattaa ttatgataca attctaatag aaaacgaatt aaattacgtt gaattgtatg
     2401 aaatctaatt gaacaagcca accacgacga cgactaacgt tgcctggatt gactcggttt
     2461 aagttaacca ctaaaaaaac ggagctgtca tgtaacacgc ggatcgagca ggtcacagtc
     2521 atgaagccat caaagcaaaa gaactaatcc aagggctgag atgattaatt agtttaaaaa
     2581 ttagttaaca cgagggaaaa ggctgtctga cagccaggtc acgttatctt tacctgtggt
     2641 cgaaatgatt cgtgtctgtc gattttaatt atttttttga aaggccgaaa ataaagttgt
     2701 aagagataaa cccgcctata taaattcata tattttcctc tccgctttga attgtctcgt
     2761 tgtcctcctc actttcatca gccgttttga atctccggcg acttgacaga gaagaacaag
     2821 gaagaagatt aagagagaaa gtaagagata atccaggaga ttcattctcc gttttgaatc
     2881 ttcctcaatc tcatcttctt ccgctctttc tttccaaggt aataggaact ttctggatct
     2941 actttatttg ctggatctcg atcttgtttt ctcaatttcc ttgagatctg gaattcgttt
     3001 aatttggatc tgtgaacctc cactaaatct tttggtttta ctagaatcga tctaagttga
     3061 ccgatcagtt agctcgatta tagctaccag aatttggctt gaccttgatg gagagatcca
     3121 tgttcatgtt acctgggaaa tgatttgtat atgtgaattg aaatctgaac tgttgaagtt
     3181 agattgaatc tgaacactgt caatgttaga ttgaatctga acactgttta aggttagatg
     3241 aagtttgtgt atagattctt cgaaacttta ggatttgtag tgtcgtacgt tgaacagaaa
     3301 gctatttctg attcaatcag ggtttatttg actgtattga actctttttg tgtgtttgca
     3361 gctcataaaa aatggctgag gctgatgata ttcaaccaat cgtgtgtgac ctcgagatga
     3421 aggataacac cgtgccactg aaattgattg ccctgttagc gaacggtgaa tttcactctg
     3481 gcgagcagtt gggtgaaacg ctgggaatga gccgggcggc tattaataaa cacattcaga
     3541 cactgcgtga ctggggcgtt gatgtcttta ccgttccggg taaaggatac agcctgcctg
     3601 agcctatcca gttacttaat gctaaacaga tattgggtca gctggatggc ggtagtgtag
     3661 ccgtgctgcc agtgattgac tccacgaatc agtaccttct tgatcgtatc ggagagctta
     3721 aatcgggcga tgcttgcatt gcagaatacc agcaggctgg ccgtggtcgc cggggtcgga
     3781 aatggttttc gccttttggc gcaaacttat atttgtcgat gttctggcgt ctggaacaag
     3841 gcccggcggc ggcgattggt ttaagtctgg ttatcggtat cgtgatggcg gaagtattac
     3901 gcaagctggg tgcagataaa gttcgtgtta aatggcctaa tgacctctat ctgcaggatc
     3961 gcaagctggc aggcattctg gtggagctga ctggcaaaac tggcgatgcg gcgcaaatag
     4021 tcattggagc cgggatcaac atggcaatgc gccgtgttga agagagtgtc gttaatcagg
     4081 ggtggatcac gctgcaggaa gcggggatca atctcgatcg taatacgttg gcggccatgc
     4141 taatacgtga attacgtgct gcgttggaac tcttcgaaca agaaggattg gcaccttatc
     4201 tgtcgcgctg ggaaaagctg gataatttta ttaatcgccc agtgaaactt atcattggtg
     4261 ataaagaaat atttggcatt tcacgcggaa tagacaaaca gggggcttta ttacttgagc
     4321 aggatggaat aataaaaccc tggatgggcg gtgaaatatc cctgcgtagt gcagaaaaat
     4381 aagctttctc cataataatg tgtgagtagt tcccagataa gggaattagg gttcctatag
     4441 ggtttcgctc atgtgttgag catataagaa acccttagta tgtatttgta tttgtaaaat
     4501 acttctatca ataaaatttc taattcctaa aaccaaaatc cagtactaaa atccagatcc
     4561 ccgtcgaccg atatccgaat tccgcttgag accacgaagt ggctcttcag tggacgaaag
     4621 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     4681 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4741 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4801 aaaaggaaga gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt
     4861 aacggcgcag tggcg
//
