LOCUS       sequence-377064-        3719 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Luz via a T7 promoter in TXTL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3719)
  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 3719)
  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-Luz
FEATURES             Location/Qualifiers
     source          1..3719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(92..111)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      905..1493
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1394..1413
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1572..1591
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1572..1590
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1599..1621
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2456..2479
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     rep_origin      2701..3156
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2788..2807)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2998..3019
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3223..3242)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3342..3364
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3402..3420)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3488..3592
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3593..3661,3662..3719,1..734)
                     /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"
ORIGIN
        1 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
       61 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
      121 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
      181 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
      241 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
      301 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
      361 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
      421 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
      481 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
      541 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tcgcgcggta
      601 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
      661 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
      721 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
      781 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
      841 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
      901 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
      961 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     1021 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     1081 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     1141 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     1201 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     1261 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     1321 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     1381 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     1441 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     1501 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atggctcgac
     1561 agatctaatt ctaatacgac tcactatagg gaaataattt tgtttaactt taagaaggag
     1621 atataccggt atgactagta tgcgtatcaa catctccctg tcgtcgttat tcgaacggtt
     1681 aagtaaactg agctctcgtt caattgcgat tacctgtgga gttgtcctcg cctctgctat
     1741 tgcttttccc atcatacgtc gggattatca gacgttcctg gaggttggtc ctagctatgc
     1801 cccgcaaaac tttcgcggct acattatcgt ttgcgtactt agcttgttcc gccaagagca
     1861 gaaaggcctg gcgatctacg accggctccc agagaagcgc cgctggctgg cggatctgcc
     1921 atttcgtgaa ggtaccagac cgagtattac ctcacacatt atccagcgcc agcgcactca
     1981 acttgtggat caggaatttg ccacacgtga actgattgac aaggttattc cgagggtgca
     2041 agcacgacat accgataaaa catttttgag cacgagcaag tttgaatttc atgcgaaagc
     2101 cattttcctg ttgccatcga ttccaatcaa cgacccgctg aacataccta gtcatgatac
     2161 ggtgcgtcga acgaaacgcg aaattgccca catgcatgac tatcacgatt gcacccttca
     2221 cctagctctg gcggcacagg atggaaaaga agtcctgaaa aaaggctggg gccagcgtca
     2281 tccgctggcg gggccgggtg tgcctgggcc gccgactgaa tggactttct tatacgcacc
     2341 ccgtaatgag gaagaagcgc gcgtggtaga aatgattgtg gaagcatcca tcggctatat
     2401 gaccaatgat ccggcaggta aaatcgtcga gaatgccaaa acgcgtggag gtggccatca
     2461 ccatcatcac catcaccacg gataactcga gcaaagcccg ccgaaaggcg ggcttttctg
     2521 tacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt
     2581 taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatccggg ctggcgtaat
     2641 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     2701 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2761 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2821 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2881 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     2941 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3001 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3061 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3121 acgcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac
     3181 gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc
     3241 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     3301 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     3361 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     3421 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     3481 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3541 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3601 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3661 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtgg
//
