LOCUS       sequence-271317-        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-014 Esp3I//(U)-AarI-SacB-AarI-(V)//...
FEATURES             Location/Qualifiers
     source          1..4630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        95..540
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(541..627,628..1962)
                     /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             2060..2719
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2126..2145)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2967..2998
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        2999..3101
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3102..3917
                     /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(3175..3194)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      3938..3965
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(3977..4564)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4057..4076)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4586..4615)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 gtccagttac gctggagtct gaggctcgtc ctgaatgcgt ctcttggaat atgcaggtgt
       61 cgcagcggac acttcgcgat cctttttaac ccatcacata tacctgccgt tcactattat
      121 ttagtgaaat gagatattat gatattttct gaattgtgat taaaaaggca actttatgcc
      181 catgcaacag aaactataaa aaatacagag aatgaaaaga aacagataga ttttttagtt
      241 ctttaggccc gtagtctgca aatcctttta tgattttcta tcaaacaaaa gaggaaaata
      301 gaccagttgc aatccaaacg agagtctaat agaatgaggt cgaaaagtaa atcgcgcggg
      361 tttgttactg ataaagcagg caagacctaa aatgtgtaaa gggcaaagtg tatactttgg
      421 cgtcacccct tacatatttt aggtcttttt ttattgtgcg taactaactt gccatcttca
      481 aacaggaggg ctggaagaag cagaccgcta acacagtaca taaaaaagga gacatgaacg
      541 atgaacatca aaaagtttgc aaaacaagca acagtattaa cctttactac cgcactgctg
      601 gcaggaggcg caactcaagc gtttgcgaaa gaaacgaacc aaaagccata taaggaaaca
      661 tacggcattt cccatattac acgccatgat atgctgcaaa tccctgaaca gcaaaaaaat
      721 gaaaaatatc aagttcctga attcgattcg tccacaatta aaaatatctc ttctgcaaaa
      781 ggcctggacg tttgggacag ctggccatta caaaacgctg acggcactgt cgcaaactat
      841 cacggctacc acatcgtctt tgcattagcc ggagatccta aaaatgcgga tgacacatcg
      901 atttacatgt tctatcaaaa agtcggcgaa acttctattg acagctggaa aaacgctggc
      961 cgcgtcttta aagacagcga caaattcgat gcaaatgatt ctatcctaaa agaccaaaca
     1021 caagaatggt caggttcagc cacatttaca tctgacggaa aaatccgttt attctacact
     1081 gatttctccg gtaaacatta cggcaaacaa acactgacaa ctgcacaagt taacgtatca
     1141 gcatcagaca gctctttgaa catcaacggt gtagaggatt ataaatcaat ctttgacggt
     1201 gacggaaaaa cgtatcaaaa tgtacagcag ttcatcgatg aaggcaacta cagctcaggc
     1261 gacaaccata cgctgagaga tcctcactac gtagaagata aaggccacaa atacttagta
     1321 tttgaagcaa acactggaac tgaagatggc taccaaggcg aagaatcttt atttaacaaa
     1381 gcatactatg gcaaaagcac atcattcttc cgtcaagaaa gtcaaaaact tctgcaaagc
     1441 gataaaaaac gcacggctga gttagcaaac ggcgctctcg gtatgattga gctaaacgat
     1501 gattacacac tgaaaaaagt gatgaaaccg ctgattgcat ctaacacagt aacagatgaa
     1561 attgaacgcg cgaacgtctt taaaatgaac ggcaaatggt acctgttcac tgactcccgc
     1621 ggatcaaaaa tgacgattga cggcattacg tctaacgata tttacatgct tggttatgtt
     1681 tctaattctt taactggccc atacaagccg ctgaacaaaa ctggccttgt gttaaaaatg
     1741 gatcttgatc ctaacgatgt aacctttact tactcacact tcgctgtacc tcaagcgaaa
     1801 ggaaacaatg tcgtgattac aagctatatg acaaacagag gattctacgc agacaaacaa
     1861 tcaacgtttg cgccaagctt cctgctgaac atcaaaggca agaaaacatc tgttgtcaaa
     1921 gacagcatcc ttgaacaagg acaattaaca gttaacaaat aaaaacgcaa aagaaaatgc
     1981 cgatatccta ttggcatttt cttttatttc ttatcaacat aaaggtgaat cccatagggc
     2041 aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc
     2101 ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa
     2161 ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa
     2221 gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaattccg
     2281 tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt
     2341 tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg
     2401 gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt
     2461 ccctaaaggg tttattgaga atatgttttt cgtgtcagcc aatccctggg tgagtttcac
     2521 cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg
     2581 caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc
     2641 cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga
     2701 gtggcagggc ggggcgtaat tcccggccag gacccaacgc tgcccgagat gcgccgcgtg
     2761 cggctgctgg agatggcgga cgcgatggat atgttctgcc aagggttggt ttgcgcattc
     2821 acagttctcc gcaagaattg attggctcca attcttggag tggtgaatcc gttagcgagg
     2881 tgccgccggc ttccatgctc ggaaagccgc acgacacctg catatgttga gagacgttta
     2941 aatcatacct gacctccata gcagaaagtc aaaagcctcc gaccggaggc ttttgacttg
     3001 atcggcacgt aagaggttcc aactttcacc ataatgaaat aagatcacta ccgggcgtat
     3061 tttttgagtt atcgagattt tcaggagcta aggaagctaa aatgagccat attcaacggg
     3121 aaacgtcttg ctcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata
     3181 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc
     3241 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag
     3301 atgagatggt caggctaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt
     3361 ttatccgtac tcctgatgat gcatggttac tcaccactgc gatcccaggg aaaacagcat
     3421 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt
     3481 tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacggc gatcgcgtat
     3541 ttcgcctcgc tcaggcgcaa tcacgaatga ataacggttt ggttggtgcg agtgattttg
     3601 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat aagcttttgc
     3661 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg
     3721 acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc
     3781 aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc
     3841 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt cacttgatgc
     3901 tcgatgagtt tttctgaggg cccaaatgta atcacctggc tcaccttcgg gtgggccttt
     3961 ctgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgat
     4021 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4081 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4141 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4201 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4261 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4321 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4381 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     4441 tgctgaagcc agttacctcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4501 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4561 tcaagaagat cctttgattt tctaccgaag aaaggcccac ccgtgaaggt gagccagtga
     4621 gttgattgca
//
