LOCUS       sequence-249596-        4617 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 4617)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 4617)
  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-010 BsaI//(A)-Esp3I-SacB-Esp3I-(Z)/...
FEATURES             Location/Qualifiers
     source          1..4617
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      12..39
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(51..638)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(131..150)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(660..689)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     promoter        791..1236
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(1237..1323,1324..2658)
                     /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             2756..3415
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2822..2841)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3655..3686
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        3687..3789
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3790..4605
                     /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(3863..3882)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 tgtaatcacc tggctcacct tcgggtgggc ctttctgcgt tgctggcgtt tttccatagg
       61 ctccgccccc ctgacgagca tcacaaaaat cgatgctcaa gtcagaggtg gcgaaacccg
      121 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
      181 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
      241 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
      301 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
      361 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
      421 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
      481 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac ctcggaaaaa
      541 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
      601 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg attttctacc
      661 gaagaaaggc ccacccgtga aggtgagcca gtgagttgat tgcagtccag ttacgctgga
      721 gtctgaggct cgtcctgaat gggtctctag agacgtcgca gcggacactt cgcgatcctt
      781 tttaacccat cacatatacc tgccgttcac tattatttag tgaaatgaga tattatgata
      841 ttttctgaat tgtgattaaa aaggcaactt tatgcccatg caacagaaac tataaaaaat
      901 acagagaatg aaaagaaaca gatagatttt ttagttcttt aggcccgtag tctgcaaatc
      961 cttttatgat tttctatcaa acaaaagagg aaaatagacc agttgcaatc caaacgagag
     1021 tctaatagaa tgaggtcgaa aagtaaatcg cgcgggtttg ttactgataa agcaggcaag
     1081 acctaaaatg tgtaaagggc aaagtgtata ctttggcgtc accccttaca tattttaggt
     1141 ctttttttat tgtgcgtaac taacttgcca tcttcaaaca ggagggctgg aagaagcaga
     1201 ccgctaacac agtacataaa aaaggagaca tgaacgatga acatcaaaaa gtttgcaaaa
     1261 caagcaacag tattaacctt tactaccgca ctgctggcag gaggcgcaac tcaagcgttt
     1321 gcgaaagaaa cgaaccaaaa gccatataag gaaacatacg gcatttccca tattacacgc
     1381 catgatatgc tgcaaatccc tgaacagcaa aaaaatgaaa aatatcaagt tcctgaattc
     1441 gattcgtcca caattaaaaa tatctcttct gcaaaaggcc tggacgtttg ggacagctgg
     1501 ccattacaaa acgctgacgg cactgtcgca aactatcacg gctaccacat cgtctttgca
     1561 ttagccggag atcctaaaaa tgcggatgac acatcgattt acatgttcta tcaaaaagtc
     1621 ggcgaaactt ctattgacag ctggaaaaac gctggccgcg tctttaaaga cagcgacaaa
     1681 ttcgatgcaa atgattctat cctaaaagac caaacacaag aatggtcagg ttcagccaca
     1741 tttacatctg acggaaaaat ccgtttattc tacactgatt tctccggtaa acattacggc
     1801 aaacaaacac tgacaactgc acaagttaac gtatcagcat cagacagctc tttgaacatc
     1861 aacggtgtag aggattataa atcaatcttt gacggtgacg gaaaaacgta tcaaaatgta
     1921 cagcagttca tcgatgaagg caactacagc tcaggcgaca accatacgct gagagatcct
     1981 cactacgtag aagataaagg ccacaaatac ttagtatttg aagcaaacac tggaactgaa
     2041 gatggctacc aaggcgaaga atctttattt aacaaagcat actatggcaa aagcacatca
     2101 ttcttccgtc aagaaagtca aaaacttctg caaagcgata aaaaacgcac ggctgagtta
     2161 gcaaacggcg ctctcggtat gattgagcta aacgatgatt acacactgaa aaaagtgatg
     2221 aaaccgctga ttgcatctaa cacagtaaca gatgaaattg aacgcgcgaa cgtctttaaa
     2281 atgaacggca aatggtacct gttcactgac tcccgcggat caaaaatgac gattgacggc
     2341 attacgtcta acgatattta catgcttggt tatgtttcta attctttaac tggcccatac
     2401 aagccgctga acaaaactgg ccttgtgtta aaaatggatc ttgatcctaa cgatgtaacc
     2461 tttacttact cacacttcgc tgtacctcaa gcgaaaggaa acaatgtcgt gattacaagc
     2521 tatatgacaa acagaggatt ctacgcagac aaacaatcaa cgtttgcgcc aagcttcctg
     2581 ctgaacatca aaggcaagaa aacatctgtt gtcaaagaca gcatccttga acaaggacaa
     2641 ttaacagtta acaaataaaa acgcaaaaga aaatgccgat atcctattgg cattttcttt
     2701 tatttcttat caacataaag gtgaatccca tagggcagga gctaaggaag ctaaaatgga
     2761 gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt
     2821 tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac
     2881 ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat
     2941 tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct
     3001 ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt
     3061 ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca
     3121 agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat
     3181 gtttttcgtg tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa
     3241 tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa
     3301 ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg
     3361 cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaattccc
     3421 ggccaggacc caacgctgcc cgagatgcgc cgcgtgcggc tgctggagat ggcggacgcg
     3481 atggatatgt tctgccaagg gttggtttgc gcattcacag ttctccgcaa gaattgattg
     3541 gctccaattc ttggagtggt gaatccgtta gcgaggtgcc gccggcttcc atgctcggaa
     3601 agccgcacga cgtctctaga gacctttaaa tcatacctga cctccatagc agaaagtcaa
     3661 aagcctccga ccggaggctt ttgacttgat cggcacgtaa gaggttccaa ctttcaccat
     3721 aatgaaataa gatcactacc gggcgtattt tttgagttat cgagattttc aggagctaag
     3781 gaagctaaaa tgagccatat tcaacgggaa acgtcttgct cgaggccgcg attaaattcc
     3841 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     3901 gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     3961 aaaggtagcg ttgccaatga tgttacagat gagatggtca ggctaaactg gctgacggaa
     4021 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     4081 accactgcga tcccagggaa aacagcattc caggtattag aagaatatcc tgattcaggt
     4141 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     4201 aattgtcctt ttaacggcga tcgcgtattt cgcctcgctc aggcgcaatc acgaatgaat
     4261 aacggtttgg ttggtgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     4321 gtctggaaag aaatgcataa gcttttgcca ttctcaccgg attcagtcgt cactcatggt
     4381 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     4441 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     4501 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     4561 atgaataaat tgcagtttca cttgatgctc gatgagtttt tctaatgagg gcccaaa
//
