LOCUS       sequence-419342-        3172 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector to store Level 0* parts.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3172)
  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 3172)
  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: pLP4
FEATURES             Location/Qualifiers
     source          1..3172
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             163..174
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             181..894
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      906..977
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      993..1020
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      1048..1105
                     /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(1129..1146)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1300..1888)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1380..1399)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2071..2165)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2191..2982,2983..3051))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2814..2833
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3052..3156)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgtctcgcga gaccgaaagt gaaacgtgat ttcatgcgtc attttgaaca ttttgtaaat
       61 cttatttaat aatgtgtgcg gcaattcaca tttaatttat gaatgttttc ttaacatcgc
      121 ggcaactcaa gaaacggcag gttcggatct tagctactag agaaagagga gaaatactag
      181 atgcgtaaag gcgaagagct gttcactggt gtcgtcccta ttctggtgga actggatggt
      241 gatgtcaacg gtcataagtt ttccgtgcgt ggcgagggtg aaggtgacgc aactaatggt
      301 aaactgacgc tgaagttcat ctgtactact ggtaaactgc cggttccttg gccgactctg
      361 gtaacgacgc tgacttatgg tgttcagtgc tttgctcgtt atccggacca tatgaagcag
      421 catgacttct tcaagtccgc catgccggaa ggctatgtgc aggaacgcac gatttccttt
      481 aaggatgacg gcacgtacaa aacgcgtgcg gaagtgaaat ttgaaggcga taccctggta
      541 aaccgcattg agctgaaagg cattgacttt aaagaggacg gcaatatcct gggccataag
      601 ctggaataca attttaacag ccacaatgtt tacatcaccg ccgataaaca aaaaaatggc
      661 attaaagcga attttaaaat tcgccacaac gtggaggatg gcagcgtgca gctggctgat
      721 cactaccagc aaaacactcc aatcggtgat ggtcctgttc tgctgccaga caatcactat
      781 ctgagcacgc aaagcgttct gtctaaagat ccgaacgaga aacgcgatca tatggttctg
      841 ctggagttcg taaccgcagc gggcatcacg catggtatgg atgaactgta caaatgacca
      901 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
      961 tgtcggtgaa cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg
     1021 tttataggtc tcatgagacg gatccagtcc ggcaaaaaag ggcaaggtgt caccaccctg
     1081 ccctttttct ttaaaaccga aaagattact tcgcgttatg caggcttcct cgctcactga
     1141 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     1201 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     1261 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccacaggc tccgcccccc
     1321 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1381 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1441 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1501 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1561 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1621 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1681 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1741 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1801 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1861 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     1921 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     1981 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2041 gtaaacttgg tctgacagct cgaggcttgg attctcacca ataaaaaacg cccggcggca
     2101 accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggat ctatcaacag
     2161 gagtccaaac gtgtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2221 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2281 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagaaccacg
     2341 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2401 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2461 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt
     2521 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     2581 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     2641 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     2701 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     2761 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac gggataatac
     2821 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     2881 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     2941 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     3001 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3061 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3121 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aa
//
