LOCUS       sequence-249557-        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-004 Esp3I//(A)-AarI-SacB-AarI-(Z)//...
FEATURES             Location/Qualifiers
     source          1..4678
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        80..525
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(526..612,613..1947)
                     /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             2045..2704
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2111..2130)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2974..3003
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3025..3612
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3513..3532
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3624..3651)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(3675..4466,4467..4535))
                     /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     4298..4317
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4536..4638)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4639..4670)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 tggaggtcag gtatgattta aacgtctctt aggatatgca ggtgtcgcag cggacacttc
       61 gcgatccttt ttaacccatc acatatacct gccgttcact attatttagt gaaatgagat
      121 attatgatat tttctgaatt gtgattaaaa aggcaacttt atgcccatgc aacagaaact
      181 ataaaaaata cagagaatga aaagaaacag atagattttt tagttcttta ggcccgtagt
      241 ctgcaaatcc ttttatgatt ttctatcaaa caaaagagga aaatagacca gttgcaatcc
      301 aaacgagagt ctaatagaat gaggtcgaaa agtaaatcgc gcgggtttgt tactgataaa
      361 gcaggcaaga cctaaaatgt gtaaagggca aagtgtatac tttggcgtca ccccttacat
      421 attttaggtc tttttttatt gtgcgtaact aacttgccat cttcaaacag gagggctgga
      481 agaagcagac cgctaacaca gtacataaaa aaggagacat gaacgatgaa catcaaaaag
      541 tttgcaaaac aagcaacagt attaaccttt actaccgcac tgctggcagg aggcgcaact
      601 caagcgtttg cgaaagaaac gaaccaaaag ccatataagg aaacatacgg catttcccat
      661 attacacgcc atgatatgct gcaaatccct gaacagcaaa aaaatgaaaa atatcaagtt
      721 cctgaattcg attcgtccac aattaaaaat atctcttctg caaaaggcct ggacgtttgg
      781 gacagctggc cattacaaaa cgctgacggc actgtcgcaa actatcacgg ctaccacatc
      841 gtctttgcat tagccggaga tcctaaaaat gcggatgaca catcgattta catgttctat
      901 caaaaagtcg gcgaaacttc tattgacagc tggaaaaacg ctggccgcgt ctttaaagac
      961 agcgacaaat tcgatgcaaa tgattctatc ctaaaagacc aaacacaaga atggtcaggt
     1021 tcagccacat ttacatctga cggaaaaatc cgtttattct acactgattt ctccggtaaa
     1081 cattacggca aacaaacact gacaactgca caagttaacg tatcagcatc agacagctct
     1141 ttgaacatca acggtgtaga ggattataaa tcaatctttg acggtgacgg aaaaacgtat
     1201 caaaatgtac agcagttcat cgatgaaggc aactacagct caggcgacaa ccatacgctg
     1261 agagatcctc actacgtaga agataaaggc cacaaatact tagtatttga agcaaacact
     1321 ggaactgaag atggctacca aggcgaagaa tctttattta acaaagcata ctatggcaaa
     1381 agcacatcat tcttccgtca agaaagtcaa aaacttctgc aaagcgataa aaaacgcacg
     1441 gctgagttag caaacggcgc tctcggtatg attgagctaa acgatgatta cacactgaaa
     1501 aaagtgatga aaccgctgat tgcatctaac acagtaacag atgaaattga acgcgcgaac
     1561 gtctttaaaa tgaacggcaa atggtacctg ttcactgact cccgcggatc aaaaatgacg
     1621 attgacggca ttacgtctaa cgatatttac atgcttggtt atgtttctaa ttctttaact
     1681 ggcccataca agccgctgaa caaaactggc cttgtgttaa aaatggatct tgatcctaac
     1741 gatgtaacct ttacttactc acacttcgct gtacctcaag cgaaaggaaa caatgtcgtg
     1801 attacaagct atatgacaaa cagaggattc tacgcagaca aacaatcaac gtttgcgcca
     1861 agcttcctgc tgaacatcaa aggcaagaaa acatctgttg tcaaagacag catccttgaa
     1921 caaggacaat taacagttaa caaataaaaa cgcaaaagaa aatgccgata tcctattggc
     1981 attttctttt atttcttatc aacataaagg tgaatcccat agggcaggag ctaaggaagc
     2041 taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa
     2101 agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct
     2161 ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt
     2221 tattcacatt cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga
     2281 cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac
     2341 tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat
     2401 atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat
     2461 tgagaatatg tttttcgtgt cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa
     2521 cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca
     2581 aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt
     2641 ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc
     2701 gtaattcccg gccaggaccc aacgctgccc gagatgcgcc gcgtgcggct gctggagatg
     2761 gcggacgcga tggatatgtt ctgccaaggg ttggtttgcg cattcacagt tctccgcaag
     2821 aattgattgg ctccaattct tggagtggtg aatccgttag cgaggtgccg ccggcttcca
     2881 tgctcggaaa gccgcacgac acctgcatat acgaagagac gcattcagga cgagcctcag
     2941 actccagcgt aactggactg caatcaactc actggctcac cttcacgggt gggcctttct
     3001 tcggtagaaa atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3061 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3121 tctttttccg aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     3181 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3241 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3301 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3361 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3421 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3481 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3541 gtcgggtttc gccacctctg acttgagcat cgatttttgt gatgctcgtc aggggggcgg
     3601 agcctatgga aaaacgccag caacgcagaa aggcccaccc gaaggtgagc caggtgatta
     3661 catttgggcc ctcattacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3721 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat gcgggagggc
     3781 ttaccatctg gccccagtgc tgcaatgata ccgcgagaac cacgctcacc ggctccagat
     3841 ttatcagcaa taaaccagcc agccgggagg gccgagcgca gaagtgatcc tgcaacttta
     3901 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     3961 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4021 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4081 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgccagaag taagttggcc
     4141 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4201 gtgagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4261 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     4321 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gtaaactctc aaggatctta
     4381 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4441 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4501 ggaataaggg cgacacggaa atgttgaata ctcattttag cttccttagc tcctgaaaat
     4561 ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa
     4621 cctcttacgt gccgatcaag tcaaaagcct ccggtcggag gcttttgact ttctgcta
//
