LOCUS       sequence-374553-        3857 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of LuxB via a T7 promoter in TXTL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3857)
  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 3857)
  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-LuxB
FEATURES             Location/Qualifiers
     source          1..3857
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      710..1298
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1199..1218
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1377..1396
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1377..1395
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1404..1426
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1445..2428
                     /codon_start=1
                     /gene="luxB"
                     /product="LuxB luciferase subunit"
                     /label=LuxB
                     /note="from the Photorhabdus luminescens bioluminescence 
                     operon"
                     /translation="MKFGLFFLNFINSTTVQEQSIVRMQEITEYVDKLNFEQILVYENH
                     FSDNGVVGAPLTVSGFLLGLTEKIKIGSLNHIITTHHPVRIAEEACLLDQLSEGRFILG
                     FSDCEKKDEMHFFNRPVEYQQQLFEECYEIINDALTTGYCNPDNDFYSFPKISVNPHAY
                     TPGGPRKYVTATSHHIVEWAAKKGIPLIFKWDDSNDVRYEYAERYKAVADKYDVDLSEI
                     DHQLMILVNYNEDSNKAKQETRAFISDYVLEMHPNENFENKLEEIIAENAVGNYTECIT
                     AAKLAIEKCGAKSVLLSFEPMNDLMSQKNVINIVDDNIKKYHMEYT"
     rep_origin      2644..3099
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2731..2750)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2941..2962
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3166..3185)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3285..3307
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3345..3363)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3431..3535
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3536..3604,3605..3857,1..539)
                     /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(3754..3773)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
       61 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
      121 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
      181 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
      241 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
      301 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
      361 ttattgctga taaatctgga gccggtgagc gtgggtcgcg cggtatcatt gcagcactgg
      421 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
      481 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
      541 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
      601 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
      661 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
      721 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
      781 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
      841 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
      901 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
      961 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     1021 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     1081 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     1141 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     1201 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     1261 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     1321 tacggttcct ggccttttgc tggccttttg ctcacatggc tcgacagatc taattctaat
     1381 acgactcact atagggaaat aattttgttt aactttaaga aggagatata ccggtatgac
     1441 tagtatgaaa tttggattgt tcttccttaa cttcatcaat tcaacaactg ttcaagaaca
     1501 aagtatagtt cgcatgcagg aaataacgga gtatgttgat aagttgaatt ttgaacagat
     1561 tttagtgtat gaaaatcatt tttcagataa tggtgttgtc ggcgctcctc tgactgtttc
     1621 tggttttctg ctcggtttaa cagagaaaat taaaattggt tcattaaatc acatcattac
     1681 aactcatcat cctgtccgca tagcggagga agcttgctta ttggatcagt taagtgaagg
     1741 gagatttatt ttagggttta gtgattgcga aaaaaaagat gaaatgcatt tttttaatcg
     1801 cccggttgaa tatcaacagc aactatttga agagtgttat gaaatcatta acgatgcttt
     1861 aacaacaggc tattgtaatc cagataacga tttttatagc ttccctaaaa tatctgtaaa
     1921 tccccatgct tatacgccag gcggacctcg gaaatatgta acagcaacca gtcatcatat
     1981 tgttgagtgg gcggccaaaa aaggtattcc tctcatcttt aagtgggatg attctaatga
     2041 tgttagatat gaatatgctg aaagatataa agccgttgcg gataaatatg acgttgacct
     2101 atcagagata gaccatcagt taatgatatt agttaactat aacgaagata gtaataaagc
     2161 taaacaagag acgcgtgcat ttattagtga ttatgttctt gaaatgcacc ctaatgaaaa
     2221 tttcgaaaat aaacttgaag aaataattgc agaaaacgct gtcggaaatt atacggagtg
     2281 tataactgcg gctaagttgg caattgaaaa gtgtggtgcg aaaagtgtat tgctgtcctt
     2341 tgaaccaatg aatgatttga tgagccaaaa aaatgtaatc aatattgttg atgataatat
     2401 taagaagtac cacatggaat atacctaact cgagcaaagc ccgccgaaag gcgggctttt
     2461 ctgtacaaca acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt
     2521 ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc gggctggcgt
     2581 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     2641 tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2701 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2761 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2821 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     2881 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     2941 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     3001 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3061 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct
     3121 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
     3181 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     3241 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     3301 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     3361 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     3421 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3481 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     3541 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3601 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     3661 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3721 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     3781 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3841 gtactcacca gtcacag
//
