LOCUS       sequence-249556-        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-003 AarI//(A)-Esp3I-SacB-Esp3I-(Z)/...
FEATURES             Location/Qualifiers
     source          1..4678
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(13..804,805..873))
                     /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     636..655
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(874..976)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(977..1008)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1096..1541
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(1542..1628,1629..2963)
                     /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             3061..3720
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3127..3146)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3990..4019
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4041..4628
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4529..4548
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4640..4667)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 tttgggccct cattaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
       61 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatgc gggagggctt
      121 accatctggc cccagtgctg caatgatacc gcgagaacca cgctcaccgg ctccagattt
      181 atcagcaata aaccagccag ccgggagggc cgagcgcaga agtgatcctg caactttatc
      241 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
      301 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
      361 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
      421 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gccagaagta agttggccgc
      481 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
      541 gagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
      601 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
      661 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcgt aaactctcaa ggatcttacc
      721 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
      781 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
      841 aataagggcg acacggaaat gttgaatact cattttagct tccttagctc ctgaaaatct
      901 cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc
      961 tcttacgtgc cgatcaagtc aaaagcctcc ggtcggaggc ttttgacttt ctgctatgga
     1021 ggtcaggtat gatttaaaca cctgcatatt aggagagacg tcgcagcgga cacttcgcga
     1081 tcctttttaa cccatcacat atacctgccg ttcactatta tttagtgaaa tgagatatta
     1141 tgatattttc tgaattgtga ttaaaaaggc aactttatgc ccatgcaaca gaaactataa
     1201 aaaatacaga gaatgaaaag aaacagatag attttttagt tctttaggcc cgtagtctgc
     1261 aaatcctttt atgattttct atcaaacaaa agaggaaaat agaccagttg caatccaaac
     1321 gagagtctaa tagaatgagg tcgaaaagta aatcgcgcgg gtttgttact gataaagcag
     1381 gcaagaccta aaatgtgtaa agggcaaagt gtatactttg gcgtcacccc ttacatattt
     1441 taggtctttt tttattgtgc gtaactaact tgccatcttc aaacaggagg gctggaagaa
     1501 gcagaccgct aacacagtac ataaaaaagg agacatgaac gatgaacatc aaaaagtttg
     1561 caaaacaagc aacagtatta acctttacta ccgcactgct ggcaggaggc gcaactcaag
     1621 cgtttgcgaa agaaacgaac caaaagccat ataaggaaac atacggcatt tcccatatta
     1681 cacgccatga tatgctgcaa atccctgaac agcaaaaaaa tgaaaaatat caagttcctg
     1741 aattcgattc gtccacaatt aaaaatatct cttctgcaaa aggcctggac gtttgggaca
     1801 gctggccatt acaaaacgct gacggcactg tcgcaaacta tcacggctac cacatcgtct
     1861 ttgcattagc cggagatcct aaaaatgcgg atgacacatc gatttacatg ttctatcaaa
     1921 aagtcggcga aacttctatt gacagctgga aaaacgctgg ccgcgtcttt aaagacagcg
     1981 acaaattcga tgcaaatgat tctatcctaa aagaccaaac acaagaatgg tcaggttcag
     2041 ccacatttac atctgacgga aaaatccgtt tattctacac tgatttctcc ggtaaacatt
     2101 acggcaaaca aacactgaca actgcacaag ttaacgtatc agcatcagac agctctttga
     2161 acatcaacgg tgtagaggat tataaatcaa tctttgacgg tgacggaaaa acgtatcaaa
     2221 atgtacagca gttcatcgat gaaggcaact acagctcagg cgacaaccat acgctgagag
     2281 atcctcacta cgtagaagat aaaggccaca aatacttagt atttgaagca aacactggaa
     2341 ctgaagatgg ctaccaaggc gaagaatctt tatttaacaa agcatactat ggcaaaagca
     2401 catcattctt ccgtcaagaa agtcaaaaac ttctgcaaag cgataaaaaa cgcacggctg
     2461 agttagcaaa cggcgctctc ggtatgattg agctaaacga tgattacaca ctgaaaaaag
     2521 tgatgaaacc gctgattgca tctaacacag taacagatga aattgaacgc gcgaacgtct
     2581 ttaaaatgaa cggcaaatgg tacctgttca ctgactcccg cggatcaaaa atgacgattg
     2641 acggcattac gtctaacgat atttacatgc ttggttatgt ttctaattct ttaactggcc
     2701 catacaagcc gctgaacaaa actggccttg tgttaaaaat ggatcttgat cctaacgatg
     2761 taacctttac ttactcacac ttcgctgtac ctcaagcgaa aggaaacaat gtcgtgatta
     2821 caagctatat gacaaacaga ggattctacg cagacaaaca atcaacgttt gcgccaagct
     2881 tcctgctgaa catcaaaggc aagaaaacat ctgttgtcaa agacagcatc cttgaacaag
     2941 gacaattaac agttaacaaa taaaaacgca aaagaaaatg ccgatatcct attggcattt
     3001 tcttttattt cttatcaaca taaaggtgaa tcccataggg caggagctaa ggaagctaaa
     3061 atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa
     3121 cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat
     3181 attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt
     3241 cacattcttg cccgcctgat gaatgctcat ccggaattcc gtatggcaat gaaagacggt
     3301 gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa
     3361 acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat
     3421 tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag
     3481 aatatgtttt tcgtgtcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg
     3541 gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc
     3601 gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat
     3661 gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa
     3721 ttcccggcca ggacccaacg ctgcccgaga tgcgccgcgt gcggctgctg gagatggcgg
     3781 acgcgatgga tatgttctgc caagggttgg tttgcgcatt cacagttctc cgcaagaatt
     3841 gattggctcc aattcttgga gtggtgaatc cgttagcgag gtgccgccgg cttccatgct
     3901 cggaaagccg cacgacgtct caacgaatat gcaggtgcat tcaggacgag cctcagactc
     3961 cagcgtaact ggactgcaat caactcactg gctcaccttc acgggtgggc ctttcttcgg
     4021 tagaaaatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4081 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4141 tttccgaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     4201 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4261 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4321 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4381 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4441 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4501 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4561 ggtttcgcca cctctgactt gagcatcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4621 tatggaaaaa cgccagcaac gcagaaaggc ccacccgaag gtgagccagg tgattaca
//
