LOCUS       sequence-383148-        3797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of LuxD via a T7 promoter in TXTL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3797)
  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 3797)
  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-LuxD
FEATURES             Location/Qualifiers
     source          1..3797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      657..1245
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1146..1165
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1324..1343
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1324..1342
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1351..1373
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1392..2315
                     /codon_start=1
                     /gene="luxD"
                     /product="LuxD acyltransferase"
                     /label=LuxD
                     /note="from the Photorhabdus luminescens bioluminescence 
                     operon"
                     /translation="MENESKYKTIDHVICVEGNKKIHVWETLPEENSPKRKNAIIIASG
                     FARRMDHFAGLAEYLSRNGFHVIRYDSLHHVGLSSGTIDEFTMSIGKQSLLAVVDWLTT
                     RKINNFGMLASSLSARIAYASLSEINASFLITAVGVVNLRYSLERALGFDYLSLPINEL
                     PNNLDFEGHKLGAEVFARDCLDFGWEDLASTINNMMYLDIPFIAFTANNDNWVKQDEVI
                     TLLSNIRSNRCKIYSLLGSSHDLSENLVVLRNFYQSVTKAAIAMDNDHLDIDVDITEPS
                     FEHLTIATVNERRMRIEIENQAISLS"
     rep_origin      2531..2986
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2618..2637)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2828..2849
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3053..3072)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3172..3194
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3232..3250)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3318..3422
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3423..3491,3492..3797,1..486)
                     /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(3641..3660)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
       61 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
      121 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
      181 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
      241 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
      301 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtcgcgcgg tatcattgca
      361 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
      421 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
      481 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
      541 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
      601 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
      661 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
      721 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
      781 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
      841 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
      901 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
      961 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1021 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1081 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1141 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     1201 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     1261 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatggctcg acagatctaa
     1321 ttctaatacg actcactata gggaaataat tttgtttaac tttaagaagg agatataccg
     1381 gtatgactag tatggaaaat gaatcaaaat ataaaaccat cgaccacgtt atttgtgttg
     1441 aaggaaataa aaaaattcat gtttgggaaa cgctgccaga agaaaacagc ccaaagagaa
     1501 agaatgccat tattattgcg tctggttttg cccgcaggat ggatcatttt gctggtctgg
     1561 cggaatattt atcgcggaat ggatttcatg tgatccgcta tgattcgctt caccacgttg
     1621 gattgagttc agggacaatt gatgaattta caatgtctat aggaaagcag agcttgttag
     1681 cagtggttga ttggttaact acacgaaaaa taaataactt cggtatgttg gcttcaagct
     1741 tatctgcgcg gatagcttat gcaagcctat ctgaaatcaa tgcttcgttt ttaatcaccg
     1801 cagtcggtgt tgttaactta agatattctc ttgaaagagc tttagggttt gattatctca
     1861 gtctacccat taatgaattg ccgaataatc tagattttga aggccataaa ttgggtgctg
     1921 aagtctttgc gagagattgt cttgattttg gttgggaaga tttagcttct acaattaata
     1981 acatgatgta tcttgatata ccgtttattg cttttactgc aaataacgat aattgggtca
     2041 agcaagatga agttatcaca ttgttatcaa atattcgtag taatcgatgc aagatatatt
     2101 ctttgttagg aagttcgcat gacttgagtg aaaatttagt ggtcctgcgc aatttttatc
     2161 aatcggttac gaaagccgct atcgcgatgg ataatgatca tctggatatt gatgttgata
     2221 ttactgaacc gtcatttgaa catttaacta ttgcgacagt caatgaacgc cgaatgagaa
     2281 ttgagattga aaatcaagca atttctctgt cttaactcga gcaaagcccg ccgaaaggcg
     2341 ggcttttctg tacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg
     2401 ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatccggg
     2461 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     2521 tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2581 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2641 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2701 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2761 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2821 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2881 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2941 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt
     3001 ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct
     3061 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     3121 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     3181 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     3241 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     3301 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3361 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3421 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     3481 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3541 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     3601 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     3661 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     3721 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     3781 tatgcagtgc tgccata
//
