LOCUS       sequence-419374-        3135 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Contains Level 0 Part: Promoter Dropout (sfGFP) for the construction
            of Level 1 plasmids and subsequent promoter insertion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3135)
  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 3135)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pLPP6
FEATURES             Location/Qualifiers
     source          1..3135
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     157..176
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     404..422
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(426..469)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    472..493
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     RBS             668..679
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             686..1399
                     /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      1411..1482
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1498..1525
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      1580..1637
                     /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(1661..1678)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1832..2420)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1912..1931)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2603..2697)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2718..3135,1..242))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 tcaccgtctt tcattgccat acgaaattcc ggatgagcat tcatcaggcg ggcaagaatg
       61 tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta
      121 atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa
      181 tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc
      241 attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat
      301 cttatttcat tatggtgaaa gttggaacct cttacgtgcc cgatcaactc gagtgccacc
      361 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
      421 gcagaatttc agataaaaaa aatccttagc tttcgctaag gatgatttct ggaattcgcg
      481 gccgcttcta gagcgatgag tggtctcggg agagagaccg aaagtgaaac gtgatttcat
      541 gcgtcatttt gaacattttg taaatcttat ttaataatgt gtgcggcaat tcacatttaa
      601 tttatgaatg ttttcttaac atcgcggcaa ctcaagaaac ggcaggttcg gatcttagct
      661 actagagaaa gaggagaaat actagatgcg taaaggcgaa gaacttttca ctggcgtagt
      721 accaatttta gtggagctgg acggagatgt aaatggtcat aagttttcag ttcgaggaga
      781 aggcgaagga gatgcaacta acggtaagct gacactaaag tttatctgta ccacgggtaa
      841 actgccggtc ccatggccga cactggttac tacacttact tatggtgtac aatgctttgc
      901 tcgttatcct gaccatatga agcagcacga tttcttcaaa tcggccatgc ctgaaggata
      961 cgtgcaagaa cgtacaatta gctttaaaga tgacggcacc tataaaacgc gcgccgaagt
     1021 taaattcgag ggtgacacat tggttaatcg tatagaactt aagggcattg acttcaaaga
     1081 agatggcaac atccttggcc ataaactgga atataatttt aactctcaca atgtctacat
     1141 tacggcggat aaacaaaaga atggcattaa agcgaatttc aaaatccgcc acaatgtcga
     1201 agacggctcg gttcaactgg cggaccatta tcagcagaac accccaatcg gtgatggccc
     1261 ggttctgtta cctgataatc attacctttc tactcaaagc gttttatcta aagatcctaa
     1321 cgagaagcgt gatcatatgg ttctactgga atttgttacc gcagctggta tcacgcacgg
     1381 catggatgag ctgtataaat gaccaggcat caaataaaac gaaaggctca gtcgaaagac
     1441 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag tcacactggc
     1501 tcaccttcgg gtgggccttt ctgcgtttat aggtctctaa tgtgagacca ctcatcgcga
     1561 ctagtagcgg ccgctgcagt ccggcaaaaa agggcaaggt gtcaccaccc tgcccttttt
     1621 ctttaaaacc gaaaagatta cttcgcgtta tgcaggcttc ctcgctcact gactcgctgc
     1681 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     1741 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     1801 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccacag gctccgcccc cctgacgagc
     1861 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     1921 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     1981 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2041 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2101 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2161 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2221 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2281 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2341 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2401 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     2461 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     2521 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     2581 ggtctgacag ctcgaggctt ggattctcac caataaaaaa cgcccggcgg caaccgagcg
     2641 ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa
     2701 gcgagctcga tatcaaatta cgccccgccc tgccactcgt cgcagtactg ttgtaattca
     2761 ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc
     2821 ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag
     2881 aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca gggattggct
     2941 gatacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa
     3001 cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc
     3061 cagagtgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta
     3121 tcccatatca ccagc
//
