LOCUS       sequence-374569-        3956 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of LuxA via a T7 promoter in TXTL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3956)
  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 3956)
  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-LuxA
FEATURES             Location/Qualifiers
     source          1..3956
                     /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..1186
                     /codon_start=1
                     /gene="luxA"
                     /product="LuxA luciferase subunit"
                     /label=LuxA
                     /note="from the Photorhabdus luminescens bioluminescence 
                     operon"
                     /translation="MKFGNFLLTYQPPQFSQTEVMKRLVKLGRISEECGFDTVWLLEHH
                     FTEFGLLGNPYVAAAYLLGATKKLNVGTAAIVLPTAHPVRQLEDVNLLDQMSKGRFRFG
                     ICRGLYNKDFRVFGTDMNNSRALAECWYGLIKNGMTEGYMEADNEHIKFHKVKVNPAAY
                     SRGGAPVYVVAESASTTEWAAQFGLPMILSWIINTNEKKAQLELYNEVAQEYGHDIHNI
                     DHCLSYITSVDHDSIKAKEICRKFLGHWYDSYVNATTIFDDSDQTRGYDFNKGQWRDFV
                     LKGHKDTNRRIDYSYEINPVGTPQECIDIIQKDIDATGISNICCGFEANGTVDEIIASM
                     KLFQSDVMPFLKEKQRSLLY"
     rep_origin      1402..1857
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1489..1508)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1699..1720
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1924..1943)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2043..2065
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2103..2121)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2189..2293
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2294..2362,2363..3154)
                     /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(2512..2531)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3325..3913
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3814..3833
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggccttttgc tcacatggct cgacagatct aattctaata cgactcacta tagggaaata
       61 attttgttta actttaagaa ggagatatac cggtatgact agtatgaaat ttggaaactt
      121 tttgcttaca taccaacctc cccaattttc tcaaacagag gtaatgaaac gtttggttaa
      181 attaggtcgc atctctgagg agtgtggttt tgataccgta tggttactgg agcatcattt
      241 cacggagttt ggtttgcttg gtaaccctta tgtcgctgct gcatatttac ttggcgcgac
      301 taaaaaattg aatgtaggaa ctgccgctat tgttcttccc acagcccatc cagtacgcca
      361 acttgaagat gtgaatttat tggatcaaat gtcaaaagga cgatttcggt ttggtatttg
      421 ccgagggctt tacaacaagg actttcgcgt attcggcaca gatatgaata acagtcgcgc
      481 cttagcggaa tgctggtacg ggctgataaa gaatggcatg acagagggat atatggaagc
      541 tgataatgaa catatcaagt tccataaggt aaaagtaaac cccgcggcgt atagcagagg
      601 tggcgcaccg gtttatgtgg tggctgaatc agcttcgacg actgagtggg ctgctcaatt
      661 tggcctaccg atgatattaa gttggattat aaatactaac gaaaagaaag cacaacttga
      721 gctttataat gaagtggctc aagaatatgg gcacgatatt cataatatcg accattgctt
      781 atcatatata acatctgtag atcatgactc aattaaagcg aaagagattt gccggaaatt
      841 tctggggcat tggtatgatt cttatgtgaa tgctacgact atttttgatg attcagacca
      901 aacaagaggt tatgatttca ataaagggca gtggcgtgac tttgtattaa aaggacataa
      961 agatactaat cgccgtattg attacagtta cgaaatcaat cccgtgggaa cgccgcagga
     1021 atgtattgac ataattcaaa aagacattga tgctacagga atatcaaata tttgttgtgg
     1081 atttgaagct aatggaacag tagacgaaat tattgcttcc atgaagctct tccagtctga
     1141 tgtcatgcca tttcttaaag aaaaacaacg ttcgctatta tattaactcg agcaaagccc
     1201 gccgaaaggc gggcttttct gtacaacaac aattgcattc attttatgtt tcaggttcag
     1261 ggggagatgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat
     1321 aaggatccgg gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     1381 cgcagcctga atggcgaatg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     1441 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     1501 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     1561 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     1621 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     1681 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     1741 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     1801 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcct
     1861 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
     1921 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     1981 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     2041 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     2101 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     2161 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     2221 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2281 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     2341 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     2401 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     2461 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     2521 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     2581 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     2641 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     2701 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     2761 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     2821 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     2881 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     2941 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     3001 tgagcgtggg tcgcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     3061 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     3121 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     3181 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     3241 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     3301 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3361 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3421 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     3481 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3541 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3601 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3661 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3721 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3781 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3841 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3901 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgct
//
