LOCUS       sequence-249624-        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-015   AarI//(U)-BsaI-SacB-BsaI-(V)/...
FEATURES             Location/Qualifiers
     source          1..4633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        97..542
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(543..629,630..1964)
                     /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             2062..2721
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2128..2147)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2969..3000
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        3001..3103
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3104..3919
                     /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(3177..3196)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      3943..3970
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(3982..4569)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4062..4081)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4591..4620)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 cagtccagtt acgctggagt ctgaggctcg tcctgaatgc acctgcatat tggaagagac
       61 ctcgcagcgg acacttcgcg atccttttta acccatcaca tatacctgcc gttcactatt
      121 atttagtgaa atgagatatt atgatatttt ctgaattgtg attaaaaagg caactttatg
      181 cccatgcaac agaaactata aaaaatacag agaatgaaaa gaaacagata gattttttag
      241 ttctttaggc ccgtagtctg caaatccttt tatgattttc tatcaaacaa aagaggaaaa
      301 tagaccagtt gcaatccaaa cgagagtcta atagaatgag gtcgaaaagt aaatcgcgcg
      361 ggtttgttac tgataaagca ggcaagacct aaaatgtgta aagggcaaag tgtatacttt
      421 ggcgtcaccc cttacatatt ttaggtcttt ttttattgtg cgtaactaac ttgccatctt
      481 caaacaggag ggctggaaga agcagaccgc taacacagta cataaaaaag gagacatgaa
      541 cgatgaacat caaaaagttt gcaaaacaag caacagtatt aacctttact accgcactgc
      601 tggcaggagg cgcaactcaa gcgtttgcga aagaaacgaa ccaaaagcca tataaggaaa
      661 catacggcat ttcccatatt acacgccatg atatgctgca aatccctgaa cagcaaaaaa
      721 atgaaaaata tcaagttcct gaattcgatt cgtccacaat taaaaatatc tcttctgcaa
      781 aaggcctgga cgtttgggac agctggccat tacaaaacgc tgacggcact gtcgcaaact
      841 atcacggcta ccacatcgtc tttgcattag ccggagatcc taaaaatgcg gatgacacat
      901 cgatttacat gttctatcaa aaagtcggcg aaacttctat tgacagctgg aaaaacgctg
      961 gccgcgtctt taaagacagc gacaaattcg atgcaaatga ttctatccta aaagaccaaa
     1021 cacaagaatg gtcaggttca gccacattta catctgacgg aaaaatccgt ttattctaca
     1081 ctgatttctc cggtaaacat tacggcaaac aaacactgac aactgcacaa gttaacgtat
     1141 cagcatcaga cagctctttg aacatcaacg gtgtagagga ttataaatca atctttgacg
     1201 gtgacggaaa aacgtatcaa aatgtacagc agttcatcga tgaaggcaac tacagctcag
     1261 gcgacaacca tacgctgaga gatcctcact acgtagaaga taaaggccac aaatacttag
     1321 tatttgaagc aaacactgga actgaagatg gctaccaagg cgaagaatct ttatttaaca
     1381 aagcatacta tggcaaaagc acatcattct tccgtcaaga aagtcaaaaa cttctgcaaa
     1441 gcgataaaaa acgcacggct gagttagcaa acggcgctct cggtatgatt gagctaaacg
     1501 atgattacac actgaaaaaa gtgatgaaac cgctgattgc atctaacaca gtaacagatg
     1561 aaattgaacg cgcgaacgtc tttaaaatga acggcaaatg gtacctgttc actgactccc
     1621 gcggatcaaa aatgacgatt gacggcatta cgtctaacga tatttacatg cttggttatg
     1681 tttctaattc tttaactggc ccatacaagc cgctgaacaa aactggcctt gtgttaaaaa
     1741 tggatcttga tcctaacgat gtaaccttta cttactcaca cttcgctgta cctcaagcga
     1801 aaggaaacaa tgtcgtgatt acaagctata tgacaaacag aggattctac gcagacaaac
     1861 aatcaacgtt tgcgccaagc ttcctgctga acatcaaagg caagaaaaca tctgttgtca
     1921 aagacagcat ccttgaacaa ggacaattaa cagttaacaa ataaaaacgc aaaagaaaat
     1981 gccgatatcc tattggcatt ttcttttatt tcttatcaac ataaaggtga atcccatagg
     2041 gcaggagcta aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata
     2101 tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat
     2161 aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac
     2221 aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattc
     2281 cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc
     2341 gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc
     2401 cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat
     2461 ttccctaaag ggtttattga gaatatgttt ttcgtgtcag ccaatccctg ggtgagtttc
     2521 accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg
     2581 ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat
     2641 gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat
     2701 gagtggcagg gcggggcgta attcccggcc aggacccaac gctgcccgag atgcgccgcg
     2761 tgcggctgct ggagatggcg gacgcgatgg atatgttctg ccaagggttg gtttgcgcat
     2821 tcacagttct ccgcaagaat tgattggctc caattcttgg agtggtgaat ccgttagcga
     2881 ggtgccgccg gcttccatgc tcggaaagcc gcacgaggtc tctgttgata tgcaggtgtt
     2941 taaatcatac ctgacctcca tagcagaaag tcaaaagcct ccgaccggag gcttttgact
     3001 tgatcggcac gtaagaggtt ccaactttca ccataatgaa ataagatcac taccgggcgt
     3061 attttttgag ttatcgagat tttcaggagc taaggaagct aaaatgagcc atattcaacg
     3121 ggaaacgtct tgctcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     3181 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa
     3241 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     3301 agatgagatg gtcaggctaa actggctgac ggaatttatg cctcttccga ccatcaagca
     3361 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatcccag ggaaaacagc
     3421 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     3481 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaacg gcgatcgcgt
     3541 atttcgcctc gctcaggcgc aatcacgaat gaataacggt ttggttggtg cgagtgattt
     3601 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataagctttt
     3661 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     3721 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     3781 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     3841 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcacttgat
     3901 gctcgatgag tttttctaat gagggcccaa atgtaatcac ctggctcacc ttcgggtggg
     3961 cctttctgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4021 tcgatgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4081 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4141 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4201 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4261 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4321 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4381 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4441 cgctctgctg aagccagtta cctcggaaaa agagttggta gctcttgatc cggcaaacaa
     4501 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4561 ggatctcaag aagatccttt gattttctac cgaagaaagg cccacccgtg aaggtgagcc
     4621 agtgagttga ttg
//
