LOCUS       sequence-249559-        4678 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 4678)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 4678)
  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-006   BsaI//(A)-AarI-SacB-AarI-(Z)/...
FEATURES             Location/Qualifiers
     source          1..4678
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        73..518
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(519..605,606..1940)
                     /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             2038..2697
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2104..2123)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2967..2996
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3018..3605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3506..3525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3617..3644)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(3668..4459,4460..4528))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4291..4310
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4529..4631)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4632..4663)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 caggtatgat ttaaaggtct cataggatat gcaggtgtcg cagcggacac ttcgcgatcc
       61 tttttaaccc atcacatata cctgccgttc actattattt agtgaaatga gatattatga
      121 tattttctga attgtgatta aaaaggcaac tttatgccca tgcaacagaa actataaaaa
      181 atacagagaa tgaaaagaaa cagatagatt ttttagttct ttaggcccgt agtctgcaaa
      241 tccttttatg attttctatc aaacaaaaga ggaaaataga ccagttgcaa tccaaacgag
      301 agtctaatag aatgaggtcg aaaagtaaat cgcgcgggtt tgttactgat aaagcaggca
      361 agacctaaaa tgtgtaaagg gcaaagtgta tactttggcg tcacccctta catattttag
      421 gtcttttttt attgtgcgta actaacttgc catcttcaaa caggagggct ggaagaagca
      481 gaccgctaac acagtacata aaaaaggaga catgaacgat gaacatcaaa aagtttgcaa
      541 aacaagcaac agtattaacc tttactaccg cactgctggc aggaggcgca actcaagcgt
      601 ttgcgaaaga aacgaaccaa aagccatata aggaaacata cggcatttcc catattacac
      661 gccatgatat gctgcaaatc cctgaacagc aaaaaaatga aaaatatcaa gttcctgaat
      721 tcgattcgtc cacaattaaa aatatctctt ctgcaaaagg cctggacgtt tgggacagct
      781 ggccattaca aaacgctgac ggcactgtcg caaactatca cggctaccac atcgtctttg
      841 cattagccgg agatcctaaa aatgcggatg acacatcgat ttacatgttc tatcaaaaag
      901 tcggcgaaac ttctattgac agctggaaaa acgctggccg cgtctttaaa gacagcgaca
      961 aattcgatgc aaatgattct atcctaaaag accaaacaca agaatggtca ggttcagcca
     1021 catttacatc tgacggaaaa atccgtttat tctacactga tttctccggt aaacattacg
     1081 gcaaacaaac actgacaact gcacaagtta acgtatcagc atcagacagc tctttgaaca
     1141 tcaacggtgt agaggattat aaatcaatct ttgacggtga cggaaaaacg tatcaaaatg
     1201 tacagcagtt catcgatgaa ggcaactaca gctcaggcga caaccatacg ctgagagatc
     1261 ctcactacgt agaagataaa ggccacaaat acttagtatt tgaagcaaac actggaactg
     1321 aagatggcta ccaaggcgaa gaatctttat ttaacaaagc atactatggc aaaagcacat
     1381 cattcttccg tcaagaaagt caaaaacttc tgcaaagcga taaaaaacgc acggctgagt
     1441 tagcaaacgg cgctctcggt atgattgagc taaacgatga ttacacactg aaaaaagtga
     1501 tgaaaccgct gattgcatct aacacagtaa cagatgaaat tgaacgcgcg aacgtcttta
     1561 aaatgaacgg caaatggtac ctgttcactg actcccgcgg atcaaaaatg acgattgacg
     1621 gcattacgtc taacgatatt tacatgcttg gttatgtttc taattcttta actggcccat
     1681 acaagccgct gaacaaaact ggccttgtgt taaaaatgga tcttgatcct aacgatgtaa
     1741 cctttactta ctcacacttc gctgtacctc aagcgaaagg aaacaatgtc gtgattacaa
     1801 gctatatgac aaacagagga ttctacgcag acaaacaatc aacgtttgcg ccaagcttcc
     1861 tgctgaacat caaaggcaag aaaacatctg ttgtcaaaga cagcatcctt gaacaaggac
     1921 aattaacagt taacaaataa aaacgcaaaa gaaaatgccg atatcctatt ggcattttct
     1981 tttatttctt atcaacataa aggtgaatcc catagggcag gagctaagga agctaaaatg
     2041 gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat
     2101 tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt
     2161 acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac
     2221 attcttgccc gcctgatgaa tgctcatccg gaattccgta tggcaatgaa agacggtgag
     2281 ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg
     2341 ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca catatattcg
     2401 caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat
     2461 atgtttttcg tgtcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc
     2521 aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac
     2581 aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc
     2641 ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaattc
     2701 ccggccagga cccaacgctg cccgagatgc gccgcgtgcg gctgctggag atggcggacg
     2761 cgatggatat gttctgccaa gggttggttt gcgcattcac agttctccgc aagaattgat
     2821 tggctccaat tcttggagtg gtgaatccgt tagcgaggtg ccgccggctt ccatgctcgg
     2881 aaagccgcac gacacctgca tatacgatga gacccattca ggacgagcct cagactccag
     2941 cgtaactgga ctgcaatcaa ctcactggct caccttcacg ggtgggcctt tcttcggtag
     3001 aaaatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3061 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3121 ccgaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     3181 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3241 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3301 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3361 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3421 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3481 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3541 ttcgccacct ctgacttgag catcgatttt tgtgatgctc gtcagggggg cggagcctat
     3601 ggaaaaacgc cagcaacgca gaaaggccca cccgaaggtg agccaggtga ttacatttgg
     3661 gccctcatta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3721 catccatagt tgcctgactc cccgtcgtgt agataactac gatgcgggag ggcttaccat
     3781 ctggccccag tgctgcaatg ataccgcgag aaccacgctc accggctcca gatttatcag
     3841 caataaacca gccagccggg agggccgagc gcagaagtga tcctgcaact ttatccgcct
     3901 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3961 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4021 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4081 aaaaagcggt tagctccttc ggtcctccga tcgttgccag aagtaagttg gccgcagtgt
     4141 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtgagat
     4201 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4261 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4321 aagtgctcat cattggaaaa cgttcttcgg ggcgtaaact ctcaaggatc ttaccgctgt
     4381 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4441 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4501 gggcgacacg gaaatgttga atactcattt tagcttcctt agctcctgaa aatctcgata
     4561 actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta
     4621 cgtgccgatc aagtcaaaag cctccggtcg gaggcttttg actttctgct atggaggt
//
