LOCUS       sequence-249558-        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-005   AarI//(A)-BsaI-SacB-BsaI-(Z)/...
FEATURES             Location/Qualifiers
     source          1..4678
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(12..803,804..872))
                     /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     635..654
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(873..975)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(976..1007)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1095..1540
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(1541..1627,1628..2962)
                     /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             3060..3719
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3126..3145)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3989..4018
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4040..4627
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4528..4547
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4639..4666)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 ttgggccctc attaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
       61 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatgcg ggagggctta
      121 ccatctggcc ccagtgctgc aatgataccg cgagaaccac gctcaccggc tccagattta
      181 tcagcaataa accagccagc cgggagggcc gagcgcagaa gtgatcctgc aactttatcc
      241 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
      301 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
      361 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
      421 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg ccagaagtaa gttggccgca
      481 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgtg
      541 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
      601 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
      661 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgta aactctcaag gatcttaccg
      721 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
      781 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
      841 ataagggcga cacggaaatg ttgaatactc attttagctt ccttagctcc tgaaaatctc
      901 gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct
      961 cttacgtgcc gatcaagtca aaagcctccg gtcggaggct tttgactttc tgctatggag
     1021 gtcaggtatg atttaaacac ctgcatatta ggagagacct cgcagcggac acttcgcgat
     1081 cctttttaac ccatcacata tacctgccgt tcactattat ttagtgaaat gagatattat
     1141 gatattttct gaattgtgat taaaaaggca actttatgcc catgcaacag aaactataaa
     1201 aaatacagag aatgaaaaga aacagataga ttttttagtt ctttaggccc gtagtctgca
     1261 aatcctttta tgattttcta tcaaacaaaa gaggaaaata gaccagttgc aatccaaacg
     1321 agagtctaat agaatgaggt cgaaaagtaa atcgcgcggg tttgttactg ataaagcagg
     1381 caagacctaa aatgtgtaaa gggcaaagtg tatactttgg cgtcacccct tacatatttt
     1441 aggtcttttt ttattgtgcg taactaactt gccatcttca aacaggaggg ctggaagaag
     1501 cagaccgcta acacagtaca taaaaaagga gacatgaacg atgaacatca aaaagtttgc
     1561 aaaacaagca acagtattaa cctttactac cgcactgctg gcaggaggcg caactcaagc
     1621 gtttgcgaaa gaaacgaacc aaaagccata taaggaaaca tacggcattt cccatattac
     1681 acgccatgat atgctgcaaa tccctgaaca gcaaaaaaat gaaaaatatc aagttcctga
     1741 attcgattcg tccacaatta aaaatatctc ttctgcaaaa ggcctggacg tttgggacag
     1801 ctggccatta caaaacgctg acggcactgt cgcaaactat cacggctacc acatcgtctt
     1861 tgcattagcc ggagatccta aaaatgcgga tgacacatcg atttacatgt tctatcaaaa
     1921 agtcggcgaa acttctattg acagctggaa aaacgctggc cgcgtcttta aagacagcga
     1981 caaattcgat gcaaatgatt ctatcctaaa agaccaaaca caagaatggt caggttcagc
     2041 cacatttaca tctgacggaa aaatccgttt attctacact gatttctccg gtaaacatta
     2101 cggcaaacaa acactgacaa ctgcacaagt taacgtatca gcatcagaca gctctttgaa
     2161 catcaacggt gtagaggatt ataaatcaat ctttgacggt gacggaaaaa cgtatcaaaa
     2221 tgtacagcag ttcatcgatg aaggcaacta cagctcaggc gacaaccata cgctgagaga
     2281 tcctcactac gtagaagata aaggccacaa atacttagta tttgaagcaa acactggaac
     2341 tgaagatggc taccaaggcg aagaatcttt atttaacaaa gcatactatg gcaaaagcac
     2401 atcattcttc cgtcaagaaa gtcaaaaact tctgcaaagc gataaaaaac gcacggctga
     2461 gttagcaaac ggcgctctcg gtatgattga gctaaacgat gattacacac tgaaaaaagt
     2521 gatgaaaccg ctgattgcat ctaacacagt aacagatgaa attgaacgcg cgaacgtctt
     2581 taaaatgaac ggcaaatggt acctgttcac tgactcccgc ggatcaaaaa tgacgattga
     2641 cggcattacg tctaacgata tttacatgct tggttatgtt tctaattctt taactggccc
     2701 atacaagccg ctgaacaaaa ctggccttgt gttaaaaatg gatcttgatc ctaacgatgt
     2761 aacctttact tactcacact tcgctgtacc tcaagcgaaa ggaaacaatg tcgtgattac
     2821 aagctatatg acaaacagag gattctacgc agacaaacaa tcaacgtttg cgccaagctt
     2881 cctgctgaac atcaaaggca agaaaacatc tgttgtcaaa gacagcatcc ttgaacaagg
     2941 acaattaaca gttaacaaat aaaaacgcaa aagaaaatgc cgatatccta ttggcatttt
     3001 cttttatttc ttatcaacat aaaggtgaat cccatagggc aggagctaag gaagctaaaa
     3061 tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac
     3121 attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata
     3181 ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc
     3241 acattcttgc ccgcctgatg aatgctcatc cggaattccg tatggcaatg aaagacggtg
     3301 agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa
     3361 cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt
     3421 cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga
     3481 atatgttttt cgtgtcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg
     3541 ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg
     3601 acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg
     3661 tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaat
     3721 tcccggccag gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg agatggcgga
     3781 cgcgatggat atgttctgcc aagggttggt ttgcgcattc acagttctcc gcaagaattg
     3841 attggctcca attcttggag tggtgaatcc gttagcgagg tgccgccggc ttccatgctc
     3901 ggaaagccgc acgaggtctc aacgaatatg caggtgcatt caggacgagc ctcagactcc
     3961 agcgtaactg gactgcaatc aactcactgg ctcaccttca cgggtgggcc tttcttcggt
     4021 agaaaatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4081 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4141 ttccgaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     4201 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4261 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4321 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4381 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4441 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4501 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4561 gtttcgccac ctctgacttg agcatcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4621 atggaaaaac gccagcaacg cagaaaggcc cacccgaagg tgagccaggt gattacat
//
