LOCUS       sequence-435251-        3071 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NanoLuc reporter plasmid to optimize chloroplast cell-free systems 
            with RBS BBa_B0034 in 5'UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3071)
  AUTHORS   Bohm CV, Inckemann R, Burgis M, Baumann J, Brinkmann CK, Lipinska 
            KE, Gilles S, Freudigmann J, Seiler VN, Clark LG, Jewett MC, Voll 
            LM, Niederholtmeyer H
  TITLE     Chloroplast Cell-Free Systems from Different Plant Species as a 
            Rapid Prototyping Platform.
  JOURNAL   ACS Synth Biol. 2024 Aug 16;13(8):2412-2424. doi: 
            10.1021/acssynbio.4c00117. Epub 2024 Jul 19.
  PUBMED    39028299
REFERENCE   2  (bases 1 to 3071)
  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.4c00117"; journalName: "ACS Synth Biol"; date: 
            "2024-08-16- 16"; volume: "13"; issue: "8"; pages: "2412-2424"
COMMENT     Sequence Label: pOP22-5'UTR-BBa_B0034
FEATURES             Location/Qualifiers
     source          1..3071
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             460..471
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             complement(join(1308..2099,2100..2168))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1931..1950
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2169..2273)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2376..2964
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2865..2884
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacacgtctc gtactggacc aaaacgaaaa aaggccgctt tcgcggcctc ttttctggaa
       61 tttggtaccg agtcgacacc ctaccgtatc gctgtggcag gcacaggatc taaccggtcc
      121 tggaccggcg taagttgctg tacagccaaa tcttccgtac taattctaca ggatttgtca
      181 actgcaaacg aatttctagt cggaacggat cacgtgcacc ggggtatgcc gtggttgaat
      241 atcggcacat gaggggctcg tgggacactt ccagaactgt ctggcgtgcg aagtctctgc
      301 taaatgcaaa acagtcagga ctcgttacag ctcgtgtatg ggagctccgg gtaggccgat
      361 cgtcaattgc ggctcggtac caaattttcg aaaaaagacg ctgaaaagcg tcttttttcg
      421 ttttggtccg gagtaatacg actcactata gtactagaga aagaggagaa ataatcaatg
      481 gttttcactt tagaagactt cgtaggtgac tggcgtcaaa cagcaggtta taatttagac
      541 caagttttag aacaaggtgg tgtatcaagt ttattccaaa atttaggtgt ttctgttact
      601 ccaattcaac gtatcgtatt aagtggtgaa aatggtctta aaattgacat ccatgttatt
      661 attccttatg aaggtctttc aggtgaccaa atgggtcaaa ttgaaaaaat ttttaaagta
      721 gtttatcctg tagatgacca tcactttaaa gtaattttac actatggtac tttagtaatt
      781 gatggcgtta cacctaatat gattgactac tttggtcgtc cttatgaagg tattgctgtt
      841 tttgatggta aaaaaatcac agttacaggt acattatgga atggtaataa aattattgac
      901 gaacgtttaa tcaatcctga tggttcatta ttattccgtg ttacaattaa tggtgttaca
      961 ggttggcgtc tttgtgaacg tatccttgct gcttaaataa taaataacgg attgtgtccg
     1021 taatcacacg tggtgcgtac gataacgcat agtgtttttc cctccactta aatcgaaggg
     1081 ttgtgtcttg gatcgcgcgg gtcaaatgta tatggttcat atacatccgc aggcacgtaa
     1141 taaagcgagg ggttcgaatc cccccgttac ccccggtagg ggcccacgct tactggagac
     1201 gagctgcggc cgcgattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     1261 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     1321 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     1381 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     1441 ataccgcggg acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     1501 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     1561 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     1621 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     1681 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     1741 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     1801 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     1861 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     1921 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     1981 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     2041 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     2101 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     2161 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     2221 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     2281 ccccgaaaag tgccacctgt catgaccaaa atcccttaac gtgagttttc gttccactga
     2341 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     2401 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     2461 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     2521 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     2581 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     2641 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     2701 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     2761 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     2821 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     2881 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     2941 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     3001 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     3061 cgtgcggccg c
//
