LOCUS       sequence-370382-        3851 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Renilla luciferase via a T7 promoter in TXTL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3851)
  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 3851)
  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-RLuc
FEATURES             Location/Qualifiers
     source          1..3851
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      708..1296
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1197..1216
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1375..1394
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1375..1393
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1402..1424
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1443..2375
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             2391..2414
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     rep_origin      2636..3091
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2723..2742)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2933..2954
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3158..3177)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3277..3299
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3337..3355)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3423..3527
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3528..3596,3597..3851,1..537)
                     /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(3746..3765)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
       61 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
      121 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
      181 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
      241 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
      301 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
      361 attgctgata aatctggagc cggtgagcgt gggtcgcgcg gtatcattgc agcactgggg
      421 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
      481 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
      541 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
      601 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
      661 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
      721 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
      781 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
      841 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
      901 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
      961 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     1021 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     1081 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     1141 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     1201 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     1261 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     1321 cggttcctgg ccttttgctg gccttttgct cacatggctc gacagatcta attctaatac
     1381 gactcactat agggaaataa ttttgtttaa ctttaagaag gagatatacc ggtatgacta
     1441 gtatgacttc gaaagtttat gatccagaac aaaggaaacg gatgataact ggtccgcagt
     1501 ggtgggccag atgtaaacaa atgaatgttc ttgattcatt tattaattat tatgattcag
     1561 aaaaacatgc agaaaatgct gttatttttt tacatggtaa cgcggcctct tcttatttat
     1621 ggcgacatgt tgtgccacat attgagccag tagcgcggtg tattatacca gaccttattg
     1681 gtatgggcaa atcaggcaaa tctggtaatg gttcttatag gttacttgat cattacaaat
     1741 atcttactgc atggtttgaa cttcttaatt taccaaagaa gatcattttt gtcggccatg
     1801 attggggtgc ttgtttggca tttcattata gctatgagca tcaagataag atcaaagcaa
     1861 tagttcacgc tgaaagtgta gtagatgtga ttgaatcatg ggatgaatgg cctgatattg
     1921 aagaagatat tgcgttgatc aaatctgaag aaggagaaaa aatggttttg gagaataact
     1981 tcttcgtgga aaccatgttg ccatcaaaaa tcatgagaaa gttagaacca gaagaatttg
     2041 cagcatatct tgaaccattc aaagagaaag gtgaagttcg tcgtccaaca ttatcatggc
     2101 ctcgtgaaat cccgttagta aaaggtggta aacctgacgt tgtacaaatt gttaggaatt
     2161 ataatgctta tctacgtgca agtgatgatt taccaaaaat gtttattgaa tcggacccag
     2221 gattcttttc caatgctatt gttgaaggtg ccaagaagtt tcctaatact gaatttgtca
     2281 aagtaaaagg tcttcatttt tcgcaagaag atgcacctga tgaaatggga aaatatatca
     2341 aatcgttcgt tgagcgagtt ctcaaaaatg aacaaacgcg tggaggtggc catcaccatc
     2401 atcaccatca ccacggataa ctcgagcaaa gcccgccgaa aggcgggctt ttctgtacaa
     2461 caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag
     2521 caagtaaaac ctctacaaat gtggtaaaat cgataaggat ccgggctggc gtaatagcga
     2581 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg
     2641 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2701 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2761 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     2821 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     2881 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     2941 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3001 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3061 aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
     3121 tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat
     3181 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     3241 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     3301 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     3361 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     3421 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3481 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3541 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3601 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3661 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3721 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     3781 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     3841 cagtcacaga a
//
