LOCUS       sequence-374564-        4316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of LuxC via a T7 promoter in TXTL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4316)
  AUTHORS   Sato W, Rasmussen M, Deich C, Engelhart AE, Adamala KP
  TITLE     Expanding luciferase reporter systems for cell-free protein 
            expression.
  JOURNAL   Sci Rep. 2022 Jul 7;12(1):11489. doi: 10.1038/s41598-022-15624-6.
  PUBMED    35798760
REFERENCE   2  (bases 1 to 4316)
  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"; journalName: "Sci Rep."; 
            date: "2022-07-7"; volume: "12(1):11489. doi"; pages: " 
            10.1038/s41598-022-15624-6"
COMMENT     Sequence Label: pTXTL-T7max-LuxC
FEATURES             Location/Qualifiers
     source          1..4316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     36..55
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        36..54
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             63..85
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             104..1546
                     /codon_start=1
                     /gene="luxC"
                     /product="LuxC fatty acid reductase"
                     /label=LuxC
                     /note="from the Photorhabdus luminescens bioluminescence 
                     operon"
                     /translation="MTKKISFIINGQVEIFPEGDDLVQSINFGDNSVYLPILNDSHVKN
                     IIDCNGNNELRLHNIVNFLYTVGQRWKNEEYSRRRTYIRDLKKYMGYSEEMAKLEANWI
                     SMILCSKGGLYDVVENELGSRHIMDEWLPQDESYVRAFPKGKSVHLLAGNVPLSGIMSI
                     LRAILTKNQCIIKTSSTDPFTANALALSFIDVDPNHPITRSLSVIYWPHQGDTSLAKEI
                     MQHADVIVAWGGPDAINWAVEHAPSYADVIKFGSKKSLCIIDNPVDLTSAATGAAHDVC
                     FYDQRACFSAQNIYYMGNHYEEFKLALIEKLNLYAHILPNAKKDFDEKAAYSLVQKESL
                     FAGLKVEVDIHQRWMIIESNAGVEFNQPLGRCVYLHHVDNIEQILPYVQKNKTQTISIF
                     PWESSFKYRDALALKGAERIVEAGMNNIFRVGGSHDGMRPLQRLVTYISHERPSNYTAK
                     DVAVEIEQTRFLEEDKFLVFVP"
     rep_origin      1762..2217
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1849..1868)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2059..2080
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2284..2303)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2403..2425
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2463..2481)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2549..2653
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2654..2722,2723..3514)
                     /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     complement(2872..2891)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3685..4273
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4174..4193
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggccttttgc tcacatggct cgacagatct aattctaata cgactcacta tagggaaata
       61 attttgttta actttaagaa ggagatatac cggtatgact agtatgacta aaaaaatttc
      121 attcattatt aacggccagg ttgaaatctt tcccgaaggt gatgatttag tgcaatccat
      181 taattttggt gataatagtg tttacctgcc aatattgaat gactctcatg taaaaaacat
      241 tattgattgt aatggaaata acgaattacg gttgcataac attgtcaatt ttctctatac
      301 ggtagggcaa agatggaaaa atgaagaata ctcaagacgc aggacataca ttcgtgactt
      361 aaaaaaatat atgggatatt cagaagaaat ggctaagcta gaggccaatt ggatatctat
      421 gattttatgt tctaaaggcg gcctttatga tgttgtagaa aatgaacttg gttctcgcca
      481 tatcatggat gaatggctac ctcaggatga aagttatgtt cgggcttttc cgaaaggtaa
      541 atctgtacat ctgttggcag gtaatgttcc attatctggg atcatgtcta tattacgcgc
      601 aattttaact aagaatcagt gtattataaa aacatcgtca accgatcctt ttaccgctaa
      661 tgcattagcg ttaagtttta ttgatgtaga ccctaatcat ccgataacgc gctctttatc
      721 tgttatatat tggccccacc aaggtgatac atcactcgca aaagaaatta tgcaacatgc
      781 ggatgttatt gtcgcttggg gagggccaga tgcgattaat tgggcggtag agcatgcgcc
      841 atcttatgct gatgtgatta aatttggttc taaaaagagt ctttgcatta tcgataatcc
      901 tgttgatttg acgtccgcag cgacaggtgc ggctcatgat gtttgttttt acgatcagcg
      961 agcttgtttt tctgcccaaa acatatatta catgggaaat cattatgagg aatttaagtt
     1021 agcgttgata gaaaaactta atctatatgc gcatatatta ccgaatgcca aaaaagattt
     1081 tgatgaaaag gcggcctatt ctttagttca aaaagaaagc ttgtttgctg gattaaaagt
     1141 agaggtggat attcatcaac gttggatgat tattgagtca aatgcaggtg tggaatttaa
     1201 tcaaccactt ggcagatgtg tgtaccttca tcacgtcgat aatattgagc aaatattgcc
     1261 ttatgttcaa aaaaataaga cgcaaaccat atctattttt ccttgggagt catcatttaa
     1321 atatcgagat gcgttagcat taaaaggtgc ggaaaggatt gtagaagcag gaatgaataa
     1381 catatttcga gttggtggat ctcatgacgg aatgagaccg ttgcaacgat tagtgacata
     1441 tatttctcat gaaaggccat ctaactatac ggctaaggat gttgcggttg aaatagaaca
     1501 gactcgattc ctggaagaag ataagttcct tgtatttgtc ccataactcg agcaaagccc
     1561 gccgaaaggc gggcttttct gtacaacaac aattgcattc attttatgtt tcaggttcag
     1621 ggggagatgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat
     1681 aaggatccgg gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     1741 cgcagcctga atggcgaatg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     1801 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     1861 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     1921 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     1981 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     2041 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     2101 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     2161 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcct
     2221 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
     2281 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     2341 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     2401 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     2461 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     2521 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     2581 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2641 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     2701 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     2761 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     2821 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     2881 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     2941 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     3001 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     3061 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     3121 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     3181 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     3241 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     3301 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     3361 tgagcgtggg tcgcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     3421 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     3481 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     3541 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     3601 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     3661 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3721 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3781 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     3841 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3901 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3961 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4021 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4081 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4141 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4201 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4261 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgct
//
