LOCUS       sequence-273316-        4630 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 4630)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 4630)
  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-013 AarI//(U)-Esp3I-SacB-Esp3I-(V)/...
FEATURES             Location/Qualifiers
     source          1..4630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        93..538
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(539..625,626..1960)
                     /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             2058..2717
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2124..2143)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2965..2996
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        2997..3099
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3100..3915
                     /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(3173..3192)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      3936..3963
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(3975..4562)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4055..4074)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4584..4613)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 ccagttacgc tggagtctga ggctcgtcct gaatgcacct gcatattgga agagacgtcg
       61 cagcggacac ttcgcgatcc tttttaaccc atcacatata cctgccgttc actattattt
      121 agtgaaatga gatattatga tattttctga attgtgatta aaaaggcaac tttatgccca
      181 tgcaacagaa actataaaaa atacagagaa tgaaaagaaa cagatagatt ttttagttct
      241 ttaggcccgt agtctgcaaa tccttttatg attttctatc aaacaaaaga ggaaaataga
      301 ccagttgcaa tccaaacgag agtctaatag aatgaggtcg aaaagtaaat cgcgcgggtt
      361 tgttactgat aaagcaggca agacctaaaa tgtgtaaagg gcaaagtgta tactttggcg
      421 tcacccctta catattttag gtcttttttt attgtgcgta actaacttgc catcttcaaa
      481 caggagggct ggaagaagca gaccgctaac acagtacata aaaaaggaga catgaacgat
      541 gaacatcaaa aagtttgcaa aacaagcaac agtattaacc tttactaccg cactgctggc
      601 aggaggcgca actcaagcgt ttgcgaaaga aacgaaccaa aagccatata aggaaacata
      661 cggcatttcc catattacac gccatgatat gctgcaaatc cctgaacagc aaaaaaatga
      721 aaaatatcaa gttcctgaat tcgattcgtc cacaattaaa aatatctctt ctgcaaaagg
      781 cctggacgtt tgggacagct ggccattaca aaacgctgac ggcactgtcg caaactatca
      841 cggctaccac atcgtctttg cattagccgg agatcctaaa aatgcggatg acacatcgat
      901 ttacatgttc tatcaaaaag tcggcgaaac ttctattgac agctggaaaa acgctggccg
      961 cgtctttaaa gacagcgaca aattcgatgc aaatgattct atcctaaaag accaaacaca
     1021 agaatggtca ggttcagcca catttacatc tgacggaaaa atccgtttat tctacactga
     1081 tttctccggt aaacattacg gcaaacaaac actgacaact gcacaagtta acgtatcagc
     1141 atcagacagc tctttgaaca tcaacggtgt agaggattat aaatcaatct ttgacggtga
     1201 cggaaaaacg tatcaaaatg tacagcagtt catcgatgaa ggcaactaca gctcaggcga
     1261 caaccatacg ctgagagatc ctcactacgt agaagataaa ggccacaaat acttagtatt
     1321 tgaagcaaac actggaactg aagatggcta ccaaggcgaa gaatctttat ttaacaaagc
     1381 atactatggc aaaagcacat cattcttccg tcaagaaagt caaaaacttc tgcaaagcga
     1441 taaaaaacgc acggctgagt tagcaaacgg cgctctcggt atgattgagc taaacgatga
     1501 ttacacactg aaaaaagtga tgaaaccgct gattgcatct aacacagtaa cagatgaaat
     1561 tgaacgcgcg aacgtcttta aaatgaacgg caaatggtac ctgttcactg actcccgcgg
     1621 atcaaaaatg acgattgacg gcattacgtc taacgatatt tacatgcttg gttatgtttc
     1681 taattcttta actggcccat acaagccgct gaacaaaact ggccttgtgt taaaaatgga
     1741 tcttgatcct aacgatgtaa cctttactta ctcacacttc gctgtacctc aagcgaaagg
     1801 aaacaatgtc gtgattacaa gctatatgac aaacagagga ttctacgcag acaaacaatc
     1861 aacgtttgcg ccaagcttcc tgctgaacat caaaggcaag aaaacatctg ttgtcaaaga
     1921 cagcatcctt gaacaaggac aattaacagt taacaaataa aaacgcaaaa gaaaatgccg
     1981 atatcctatt ggcattttct tttatttctt atcaacataa aggtgaatcc catagggcag
     2041 gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc
     2101 aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc
     2161 agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt
     2221 tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg gaattccgta
     2281 tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt
     2341 tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc
     2401 agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc
     2461 ctaaagggtt tattgagaat atgtttttcg tgtcagccaa tccctgggtg agtttcacca
     2521 gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca
     2581 aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg
     2641 tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt
     2701 ggcagggcgg ggcgtaattc ccggccagga cccaacgctg cccgagatgc gccgcgtgcg
     2761 gctgctggag atggcggacg cgatggatat gttctgccaa gggttggttt gcgcattcac
     2821 agttctccgc aagaattgat tggctccaat tcttggagtg gtgaatccgt tagcgaggtg
     2881 ccgccggctt ccatgctcgg aaagccgcac gacgtctctg ttgatatgca ggtgtttaaa
     2941 tcatacctga cctccatagc agaaagtcaa aagcctccga ccggaggctt ttgacttgat
     3001 cggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt
     3061 tttgagttat cgagattttc aggagctaag gaagctaaaa tgagccatat tcaacgggaa
     3121 acgtcttgct cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
     3181 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgattgta tgggaagccc
     3241 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     3301 gagatggtca ggctaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
     3361 atccgtactc ctgatgatgc atggttactc accactgcga tcccagggaa aacagcattc
     3421 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
     3481 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacggcga tcgcgtattt
     3541 cgcctcgctc aggcgcaatc acgaatgaat aacggtttgg ttggtgcgag tgattttgat
     3601 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa gcttttgcca
     3661 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     3721 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     3781 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     3841 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca cttgatgctc
     3901 gatgagtttt tctgagggcc caaatgtaat cacctggctc accttcgggt gggcctttct
     3961 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgatgc
     4021 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4081 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4141 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4201 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4261 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4321 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4381 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4441 ctgaagccag ttacctcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4501 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4561 aagaagatcc tttgattttc taccgaagaa aggcccaccc gtgaaggtga gccagtgagt
     4621 tgattgcagt
//
