LOCUS       sequence-374550-        3986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of LuxE via a T7 promoter in TXTL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3986)
  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 3986)
  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-LuxE
FEATURES             Location/Qualifiers
     source          1..3986
                     /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..1216
                     /codon_start=1
                     /gene="luxE"
                     /product="LuxE"
                     /label=LuxE
                     /note="from the Photorhabdus luminescens bioluminescence 
                     operon"
                     /translation="MTSYVDKQEITASSEIDDLIFSSDPLVWSYDEQEKIRKKLVLDAF
                     RNHYKHCREYRHYCQAHKVDDNITEIDDIPVFPTSVFKFTRLLTSQENEIESWFTSSGT
                     NGLKSQVARDRLSIERLLGSVSYGMKYVGSWFDHQIELVNLGPDRFNAHNIWFKYVMSL
                     VELLYPTTFTVTEERIDFVKTLNSLERIKNQGKDLCLIGSPYFIYLLCHYMKDKKISFS
                     GDKSLYIITGGGWKSYEKESLKRDDFNHLLFDTFNLSDISQIRDIFNQVELNTCFFEDE
                     MQRKHVPPWVYARALDPETLKPVPDGTPGLMSYMDASATSYPAFIVTDDVGIISREYGK
                     YPGVLVEILRRVNTRTQKGCALSLTEAFDS"
     rep_origin      1432..1887
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1519..1538)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1729..1750
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1954..1973)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2073..2095
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2133..2151)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2219..2323
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2324..2392,2393..3184)
                     /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(2542..2561)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3355..3943
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3844..3863
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggccttttgc tcacatggct cgacagatct aattctaata cgactcacta tagggaaata
       61 attttgttta actttaagaa ggagatatac cggtatgact agtatgactt catatgttga
      121 taaacaagaa attacagcaa gctcagaaat tgatgatttg attttttcga gcgatccatt
      181 agtgtggtct tacgacgagc aggaaaaaat cagaaagaaa cttgtgcttg atgcatttcg
      241 taatcattat aaacattgtc gagaatatcg tcactactgt caggcacaca aagtagatga
      301 caatattacg gaaattgatg acatacctgt attcccaaca tcggttttta agtttactcg
      361 cttattaact tctcaggaaa acgagattga aagttggttt accagtagcg gcacgaatgg
      421 tttaaaaagt caggtggcgc gtgacagatt aagtattgag agactcttag gctctgtgag
      481 ttatggcatg aaatatgttg gtagttggtt tgatcatcaa atagaattag tcaatttggg
      541 accagataga tttaatgctc ataatatttg gtttaaatat gttatgagtt tggtggaatt
      601 gttatatcct acgacattta ccgtaacaga agaacgaata gattttgtta aaacattgaa
      661 tagtcttgaa cgaataaaaa atcaagggaa agatctttgt cttattggtt cgccatactt
      721 tatttattta ctctgccatt atatgaaaga taaaaaaatc tcattttctg gagataaaag
      781 cctttatatc ataaccggag gcggctggaa aagttacgaa aaagaatctc tgaaacgtga
      841 tgatttcaat catcttttat ttgatacttt caatctcagt gatattagtc agatccgaga
      901 tatatttaat caagttgaac tcaacacttg tttctttgag gatgaaatgc agcgtaaaca
      961 tgttccgccg tgggtatatg cgcgagcgct tgatcctgaa acgttgaaac ctgtacctga
     1021 tggaacgccg gggttgatga gttatatgga tgcgtcagca accagttatc cagcatttat
     1081 tgttaccgat gatgtcggga taattagcag agaatatggt aagtatcccg gcgtgctcgt
     1141 tgaaatttta cgtcgcgtca atacgaggac gcagaaaggg tgtgctttaa gcttaaccga
     1201 agcgtttgat agttaactcg agcaaagccc gccgaaaggc gggcttttct gtacaacaac
     1261 aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag
     1321 taaaacctct acaaatgtgg taaaatcgat aaggatccgg gctggcgtaa tagcgaagag
     1381 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct
     1441 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     1501 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     1561 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     1621 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     1681 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     1741 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     1801 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     1861 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     1921 cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     1981 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     2041 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     2101 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     2161 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     2221 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     2281 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     2341 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     2401 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     2461 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     2521 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     2581 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     2641 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     2701 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     2761 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     2821 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     2881 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     2941 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     3001 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tcgcgcggta tcattgcagc
     3061 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     3121 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     3181 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     3241 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     3301 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3361 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3421 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3481 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3541 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3601 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3661 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3721 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3781 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3841 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3901 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3961 ctttttacgg ttcctggcct tttgct
//
