LOCUS       sequence-351026-        4670 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 4670)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 4670)
  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-001 BsaI//(A)-Esp3I-SacB-Esp3I-(Z)/...
FEATURES             Location/Qualifiers
     source          1..4670
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        86..531
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(532..618,619..1953)
                     /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             2051..2710
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2117..2136)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2976..3005
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3027..3614
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3515..3534
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3626..3653)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(3677..4468,4469..4537))
                     /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     4300..4319
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4538..4640)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(join(4641..4670,1..2))
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 ctttctgcta tggaggtcag gtatgattta aaggtctcat aggagagacg tcgcagcgga
       61 cacttcgcga tcctttttaa cccatcacat atacctgccg ttcactatta tttagtgaaa
      121 tgagatatta tgatattttc tgaattgtga ttaaaaaggc aactttatgc ccatgcaaca
      181 gaaactataa aaaatacaga gaatgaaaag aaacagatag attttttagt tctttaggcc
      241 cgtagtctgc aaatcctttt atgattttct atcaaacaaa agaggaaaat agaccagttg
      301 caatccaaac gagagtctaa tagaatgagg tcgaaaagta aatcgcgcgg gtttgttact
      361 gataaagcag gcaagaccta aaatgtgtaa agggcaaagt gtatactttg gcgtcacccc
      421 ttacatattt taggtctttt tttattgtgc gtaactaact tgccatcttc aaacaggagg
      481 gctggaagaa gcagaccgct aacacagtac ataaaaaagg agacatgaac gatgaacatc
      541 aaaaagtttg caaaacaagc aacagtatta acctttacta ccgcactgct ggcaggaggc
      601 gcaactcaag cgtttgcgaa agaaacgaac caaaagccat ataaggaaac atacggcatt
      661 tcccatatta cacgccatga tatgctgcaa atccctgaac agcaaaaaaa tgaaaaatat
      721 caagttcctg aattcgattc gtccacaatt aaaaatatct cttctgcaaa aggcctggac
      781 gtttgggaca gctggccatt acaaaacgct gacggcactg tcgcaaacta tcacggctac
      841 cacatcgtct ttgcattagc cggagatcct aaaaatgcgg atgacacatc gatttacatg
      901 ttctatcaaa aagtcggcga aacttctatt gacagctgga aaaacgctgg ccgcgtcttt
      961 aaagacagcg acaaattcga tgcaaatgat tctatcctaa aagaccaaac acaagaatgg
     1021 tcaggttcag ccacatttac atctgacgga aaaatccgtt tattctacac tgatttctcc
     1081 ggtaaacatt acggcaaaca aacactgaca actgcacaag ttaacgtatc agcatcagac
     1141 agctctttga acatcaacgg tgtagaggat tataaatcaa tctttgacgg tgacggaaaa
     1201 acgtatcaaa atgtacagca gttcatcgat gaaggcaact acagctcagg cgacaaccat
     1261 acgctgagag atcctcacta cgtagaagat aaaggccaca aatacttagt atttgaagca
     1321 aacactggaa ctgaagatgg ctaccaaggc gaagaatctt tatttaacaa agcatactat
     1381 ggcaaaagca catcattctt ccgtcaagaa agtcaaaaac ttctgcaaag cgataaaaaa
     1441 cgcacggctg agttagcaaa cggcgctctc ggtatgattg agctaaacga tgattacaca
     1501 ctgaaaaaag tgatgaaacc gctgattgca tctaacacag taacagatga aattgaacgc
     1561 gcgaacgtct ttaaaatgaa cggcaaatgg tacctgttca ctgactcccg cggatcaaaa
     1621 atgacgattg acggcattac gtctaacgat atttacatgc ttggttatgt ttctaattct
     1681 ttaactggcc catacaagcc gctgaacaaa actggccttg tgttaaaaat ggatcttgat
     1741 cctaacgatg taacctttac ttactcacac ttcgctgtac ctcaagcgaa aggaaacaat
     1801 gtcgtgatta caagctatat gacaaacaga ggattctacg cagacaaaca atcaacgttt
     1861 gcgccaagct tcctgctgaa catcaaaggc aagaaaacat ctgttgtcaa agacagcatc
     1921 cttgaacaag gacaattaac agttaacaaa taaaaacgca aaagaaaatg ccgatatcct
     1981 attggcattt tcttttattt cttatcaaca taaaggtgaa tcccataggg caggagctaa
     2041 ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca
     2101 tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt
     2161 tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc
     2221 ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaattcc gtatggcaat
     2281 gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga
     2341 gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct
     2401 acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg
     2461 gtttattgag aatatgtttt tcgtgtcagc caatccctgg gtgagtttca ccagttttga
     2521 tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta
     2581 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga
     2641 tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg
     2701 cggggcgtaa ttcccggcca ggacccaacg ctgcccgaga tgcgccgcgt gcggctgctg
     2761 gagatggcgg acgcgatgga tatgttctgc caagggttgg tttgcgcatt cacagttctc
     2821 cgcaagaatt gattggctcc aattcttgga gtggtgaatc cgttagcgag gtgccgccgg
     2881 cttccatgct cggaaagccg cacgacgtct caacgatgag acccattcag gacgagcctc
     2941 agactccagc gtaactggac tgcaatcaac tcactggctc accttcacgg gtgggccttt
     3001 cttcggtaga aaatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3061 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3121 actctttttc cgaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     3181 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3241 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3301 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3361 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3421 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3481 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3541 ctgtcgggtt tcgccacctc tgacttgagc atcgattttt gtgatgctcg tcaggggggc
     3601 ggagcctatg gaaaaacgcc agcaacgcag aaaggcccac ccgaaggtga gccaggtgat
     3661 tacatttggg ccctcattac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3721 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atgcgggagg
     3781 gcttaccatc tggccccagt gctgcaatga taccgcgaga accacgctca ccggctccag
     3841 atttatcagc aataaaccag ccagccggga gggccgagcg cagaagtgat cctgcaactt
     3901 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     3961 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4021 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4081 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgccaga agtaagttgg
     4141 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4201 ccgtgagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4261 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4321 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgtaaactc tcaaggatct
     4381 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4441 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4501 agggaataag ggcgacacgg aaatgttgaa tactcatttt agcttcctta gctcctgaaa
     4561 atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg
     4621 aacctcttac gtgccgatca agtcaaaagc ctccggtcgg aggcttttga
//
