LOCUS       sequence-327630-        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-002 Esp3I//(A)-BsaI-SacB-BsaI-(Z)//...
FEATURES             Location/Qualifiers
     source          1..4670
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        77..522
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(523..609,610..1944)
                     /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             2042..2701
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2108..2127)
                     /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 atggaggtca ggtatgattt aaacgtctca taggagagac ctcgcagcgg acacttcgcg
       61 atccttttta acccatcaca tatacctgcc gttcactatt atttagtgaa atgagatatt
      121 atgatatttt ctgaattgtg attaaaaagg caactttatg cccatgcaac agaaactata
      181 aaaaatacag agaatgaaaa gaaacagata gattttttag ttctttaggc ccgtagtctg
      241 caaatccttt tatgattttc tatcaaacaa aagaggaaaa tagaccagtt gcaatccaaa
      301 cgagagtcta atagaatgag gtcgaaaagt aaatcgcgcg ggtttgttac tgataaagca
      361 ggcaagacct aaaatgtgta aagggcaaag tgtatacttt ggcgtcaccc cttacatatt
      421 ttaggtcttt ttttattgtg cgtaactaac ttgccatctt caaacaggag ggctggaaga
      481 agcagaccgc taacacagta cataaaaaag gagacatgaa cgatgaacat caaaaagttt
      541 gcaaaacaag caacagtatt aacctttact accgcactgc tggcaggagg cgcaactcaa
      601 gcgtttgcga aagaaacgaa ccaaaagcca tataaggaaa catacggcat ttcccatatt
      661 acacgccatg atatgctgca aatccctgaa cagcaaaaaa atgaaaaata tcaagttcct
      721 gaattcgatt cgtccacaat taaaaatatc tcttctgcaa aaggcctgga cgtttgggac
      781 agctggccat tacaaaacgc tgacggcact gtcgcaaact atcacggcta ccacatcgtc
      841 tttgcattag ccggagatcc taaaaatgcg gatgacacat cgatttacat gttctatcaa
      901 aaagtcggcg aaacttctat tgacagctgg aaaaacgctg gccgcgtctt taaagacagc
      961 gacaaattcg atgcaaatga ttctatccta aaagaccaaa cacaagaatg gtcaggttca
     1021 gccacattta catctgacgg aaaaatccgt ttattctaca ctgatttctc cggtaaacat
     1081 tacggcaaac aaacactgac aactgcacaa gttaacgtat cagcatcaga cagctctttg
     1141 aacatcaacg gtgtagagga ttataaatca atctttgacg gtgacggaaa aacgtatcaa
     1201 aatgtacagc agttcatcga tgaaggcaac tacagctcag gcgacaacca tacgctgaga
     1261 gatcctcact acgtagaaga taaaggccac aaatacttag tatttgaagc aaacactgga
     1321 actgaagatg gctaccaagg cgaagaatct ttatttaaca aagcatacta tggcaaaagc
     1381 acatcattct tccgtcaaga aagtcaaaaa cttctgcaaa gcgataaaaa acgcacggct
     1441 gagttagcaa acggcgctct cggtatgatt gagctaaacg atgattacac actgaaaaaa
     1501 gtgatgaaac cgctgattgc atctaacaca gtaacagatg aaattgaacg cgcgaacgtc
     1561 tttaaaatga acggcaaatg gtacctgttc actgactccc gcggatcaaa aatgacgatt
     1621 gacggcatta cgtctaacga tatttacatg cttggttatg tttctaattc tttaactggc
     1681 ccatacaagc cgctgaacaa aactggcctt gtgttaaaaa tggatcttga tcctaacgat
     1741 gtaaccttta cttactcaca cttcgctgta cctcaagcga aaggaaacaa tgtcgtgatt
     1801 acaagctata tgacaaacag aggattctac gcagacaaac aatcaacgtt tgcgccaagc
     1861 ttcctgctga acatcaaagg caagaaaaca tctgttgtca aagacagcat ccttgaacaa
     1921 ggacaattaa cagttaacaa ataaaaacgc aaaagaaaat gccgatatcc tattggcatt
     1981 ttcttttatt tcttatcaac ataaaggtga atcccatagg gcaggagcta aggaagctaa
     2041 aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga
     2101 acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga
     2161 tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat
     2221 tcacattctt gcccgcctga tgaatgctca tccggaattc cgtatggcaa tgaaagacgg
     2281 tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga
     2341 aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata
     2401 ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga
     2461 gaatatgttt ttcgtgtcag ccaatccctg ggtgagtttc accagttttg atttaaacgt
     2521 ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg
     2581 cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca
     2641 tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta
     2701 attcccggcc aggacccaac gctgcccgag atgcgccgcg tgcggctgct ggagatggcg
     2761 gacgcgatgg atatgttctg ccaagggttg gtttgcgcat tcacagttct ccgcaagaat
     2821 tgattggctc caattcttgg agtggtgaat ccgttagcga ggtgccgccg gcttccatgc
     2881 tcggaaagcc gcacgaggtc tcaacgatga gacgcattca 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
//
