LOCUS       sequence-419384-        3871 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains Level 0* Part: E. coli shuttle part (pMB1_cat) for the 
            construction of Level 2 plasmids.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3871)
  AUTHORS   Li X, Schonberg PY, Wucherpfennig T, Hinze C, Sulaj F, Henle T, 
            Mascher T
  TITLE     Development of a Golden Gate Assembly-Based Genetic Toolbox for 
            Lactiplantibacillus plantarum and Its Application for Engineering 
            Monoterpenoid Biosynthesis.
  JOURNAL   ACS Synth Biol. 2024 Sep 20;13(9):2764-2779. doi: 
            10.1021/acssynbio.4c00075. Epub 2024 Sep 10.
  PUBMED    39254046
REFERENCE   2  (bases 1 to 3871)
  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"; doi: 
            "10.1021/acssynbio.4c00075"; journalName: "ACS Synth Biol"; date: 
            "2024-09-20- 20"; volume: "13"; issue: "9"; pages: "2764-2779"
COMMENT     Sequence Label: pLPE3
FEATURES             Location/Qualifiers
     source          1..3871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             158..169
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      883..954
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      970..997
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      1012..1069
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(1093..1110)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1264..1852)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(2035..2129)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2277..2936
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2343..2362)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2957..3051
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      3234..3822
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tgagacgcgg aaagtgaaac gtgatttcat gcgtcatttt gaacattttg taaatcttat
       61 ttaataatgt gtgcggcaat tcacatttaa tttatgaatg ttttcttaac atcgcggcaa
      121 ctcaagaaac ggcaggttcg gatcttagct actagagaaa gaggagaaat actagatggt
      181 ttctaaaggt gaagcagtga tcaaagaatt tatgcgcttc aaagttcaca tggaaggttc
      241 tatgaacggc cacgaatttg aaattgaagg tgaaggcgaa ggtcgtccat acgaaggtac
      301 tcaaaccgca aaactgaaag ttaccaaagg tggtccactg ccattttctt gggatattct
      361 gtctccacaa tttatgtacg gttctcgtgc atttatcaaa cacccagcag atattccaga
      421 ctactacaaa caatcttttc cggaaggttt caaatgggaa cgtgttatga attttgaaga
      481 tggtggtgca gttacggtta cccaagatac ctctctggaa gatggtactc tgatctacaa
      541 agttaaactg cgtggtacta actttccacc agatggtcca gttatgcaga aaaaaaccat
      601 gggttgggaa gcatctaccg aacgtctgta cccagaagat ggcgttctga aaggtgatat
      661 caaaatggca ctgcgtctga aagatggcgg tcgttacctg gcagatttca aaaccaccta
      721 caaagcgaaa aaaccagttc aaatgccagg tgcatacaac gttgatcgta aactggatat
      781 taccagccac aacgaagatt acaccgttgt tgaacaatac gaacgttctg aaggccgtca
      841 ctctaccggt ggtatggatg aactgtacaa ataaccaggc atcaaataaa acgaaaggct
      901 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag
      961 agtcacactg gctcaccttc gggtgggcct ttctgcgttt atacgatcca gtccggcaaa
     1021 aaagggcaag gtgtcaccac cctgcccttt ttctttaaaa ccgaaaagat tacttcgcgt
     1081 tatgcaggct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1141 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1201 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1261 gtttttccac aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1321 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1381 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1441 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1501 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     1561 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1621 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     1681 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     1741 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     1801 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     1861 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     1921 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     1981 taaatcaatc taaagtatat atgagtaaac ttggtctgac agctcgaggc ttggattctc
     2041 accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga
     2101 ggtcattact ggatctatca acaggagtcc aaacgtcgtc tcgggtcctt agacgtcagg
     2161 tggcactcga gttgatcggg cacgtaagag gttccaactt tcaccataat gaaataagat
     2221 cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatgg
     2281 agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt
     2341 ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta
     2401 cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca
     2461 ttcttgcccg cctgatgaat gctcatccgg aatttcgtat ggcaatgaaa gacggtgagc
     2521 tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt
     2581 tttcatcact ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc
     2641 aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata
     2701 tgtttttcgt atcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca
     2761 atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca
     2821 aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg
     2881 gcagaatgct taatgaatta caacagtact gcgacgagtg gcagggcggg gcgtaatttg
     2941 atatcgagct cgcttggact cctgttgata gatccagtaa tgacctcaga actccatctg
     3001 gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccaagcctc
     3061 gagctgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     3121 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     3181 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3241 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3301 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3361 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     3421 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3481 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3541 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3601 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3661 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3721 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3781 cgatttttgt gatgctcgtc aggggggcgg agcctgtgga aaaacgccag caacgcggcc
     3841 tttttacggt tcctggcctt ttgctggaac a
//
