LOCUS       sequence-249625-        4633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EMMA Kit # 1000000119 extension.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4633)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 4633)
  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"
COMMENT     Sequence Label: pRDAX-EMMI-016   BsaI//(U)-AarI-SacB-AarI-(V)/...
FEATURES             Location/Qualifiers
     source          1..4633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        91..536
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(537..623,624..1958)
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     CDS             2056..2715
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2122..2141)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2963..2994
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        2995..3097
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3098..3913
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3171..3190)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      3937..3964
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(3976..4563)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4056..4075)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4585..4614)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 agttacgctg gagtctgagg ctcgtcctga atgggtctct tggaatatgc aggtgtcgca
       61 gcggacactt cgcgatcctt tttaacccat cacatatacc tgccgttcac tattatttag
      121 tgaaatgaga tattatgata ttttctgaat tgtgattaaa aaggcaactt tatgcccatg
      181 caacagaaac tataaaaaat acagagaatg aaaagaaaca gatagatttt ttagttcttt
      241 aggcccgtag tctgcaaatc cttttatgat tttctatcaa acaaaagagg aaaatagacc
      301 agttgcaatc caaacgagag tctaatagaa tgaggtcgaa aagtaaatcg cgcgggtttg
      361 ttactgataa agcaggcaag acctaaaatg tgtaaagggc aaagtgtata ctttggcgtc
      421 accccttaca tattttaggt ctttttttat tgtgcgtaac taacttgcca tcttcaaaca
      481 ggagggctgg aagaagcaga ccgctaacac agtacataaa aaaggagaca tgaacgatga
      541 acatcaaaaa gtttgcaaaa caagcaacag tattaacctt tactaccgca ctgctggcag
      601 gaggcgcaac tcaagcgttt gcgaaagaaa cgaaccaaaa gccatataag gaaacatacg
      661 gcatttccca tattacacgc catgatatgc tgcaaatccc tgaacagcaa aaaaatgaaa
      721 aatatcaagt tcctgaattc gattcgtcca caattaaaaa tatctcttct gcaaaaggcc
      781 tggacgtttg ggacagctgg ccattacaaa acgctgacgg cactgtcgca aactatcacg
      841 gctaccacat cgtctttgca ttagccggag atcctaaaaa tgcggatgac acatcgattt
      901 acatgttcta tcaaaaagtc ggcgaaactt ctattgacag ctggaaaaac gctggccgcg
      961 tctttaaaga cagcgacaaa ttcgatgcaa atgattctat cctaaaagac caaacacaag
     1021 aatggtcagg ttcagccaca tttacatctg acggaaaaat ccgtttattc tacactgatt
     1081 tctccggtaa acattacggc aaacaaacac tgacaactgc acaagttaac gtatcagcat
     1141 cagacagctc tttgaacatc aacggtgtag aggattataa atcaatcttt gacggtgacg
     1201 gaaaaacgta tcaaaatgta cagcagttca tcgatgaagg caactacagc tcaggcgaca
     1261 accatacgct gagagatcct cactacgtag aagataaagg ccacaaatac ttagtatttg
     1321 aagcaaacac tggaactgaa gatggctacc aaggcgaaga atctttattt aacaaagcat
     1381 actatggcaa aagcacatca ttcttccgtc aagaaagtca aaaacttctg caaagcgata
     1441 aaaaacgcac ggctgagtta gcaaacggcg ctctcggtat gattgagcta aacgatgatt
     1501 acacactgaa aaaagtgatg aaaccgctga ttgcatctaa cacagtaaca gatgaaattg
     1561 aacgcgcgaa cgtctttaaa atgaacggca aatggtacct gttcactgac tcccgcggat
     1621 caaaaatgac gattgacggc attacgtcta acgatattta catgcttggt tatgtttcta
     1681 attctttaac tggcccatac aagccgctga acaaaactgg ccttgtgtta aaaatggatc
     1741 ttgatcctaa cgatgtaacc tttacttact cacacttcgc tgtacctcaa gcgaaaggaa
     1801 acaatgtcgt gattacaagc tatatgacaa acagaggatt ctacgcagac aaacaatcaa
     1861 cgtttgcgcc aagcttcctg ctgaacatca aaggcaagaa aacatctgtt gtcaaagaca
     1921 gcatccttga acaaggacaa ttaacagtta acaaataaaa acgcaaaaga aaatgccgat
     1981 atcctattgg cattttcttt tatttcttat caacataaag gtgaatccca tagggcagga
     2041 gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa
     2101 tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag
     2161 accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt
     2221 tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga attccgtatg
     2281 gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc
     2341 catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag
     2401 tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct
     2461 aaagggttta ttgagaatat gtttttcgtg tcagccaatc cctgggtgag tttcaccagt
     2521 tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa
     2581 tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt
     2641 tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg
     2701 cagggcgggg cgtaattccc ggccaggacc caacgctgcc cgagatgcgc cgcgtgcggc
     2761 tgctggagat ggcggacgcg atggatatgt tctgccaagg gttggtttgc gcattcacag
     2821 ttctccgcaa gaattgattg gctccaattc ttggagtggt gaatccgtta gcgaggtgcc
     2881 gccggcttcc atgctcggaa agccgcacga cacctgcata tgttgagaga cctttaaatc
     2941 atacctgacc tccatagcag aaagtcaaaa gcctccgacc ggaggctttt gacttgatcg
     3001 gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg gcgtattttt
     3061 tgagttatcg agattttcag gagctaagga agctaaaatg agccatattc aacgggaaac
     3121 gtcttgctcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     3181 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga
     3241 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     3301 gatggtcagg ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     3361 ccgtactcct gatgatgcat ggttactcac cactgcgatc ccagggaaaa cagcattcca
     3421 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     3481 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacggcgatc gcgtatttcg
     3541 cctcgctcag gcgcaatcac gaatgaataa cggtttggtt ggtgcgagtg attttgatga
     3601 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataagc ttttgccatt
     3661 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     3721 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     3781 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     3841 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcact tgatgctcga
     3901 tgagtttttc taatgagggc ccaaatgtaa tcacctggct caccttcggg tgggcctttc
     3961 tgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgatg
     4021 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4081 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4141 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4201 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4261 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4321 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4381 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4441 gctgaagcca gttacctcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4501 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4561 caagaagatc ctttgatttt ctaccgaaga aaggcccacc cgtgaaggtg agccagtgag
     4621 ttgattgcag tcc
//
