LOCUS       sequence-374551-        4504 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of eGFP-Luz via a T7 promoter in TXTL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4504)
  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 4504)
  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-HisGFPLuz
FEATURES             Location/Qualifiers
     source          1..4504
                     /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             101..124
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(170..191)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(431..450)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     778..799
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1678..1698
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     rep_origin      1950..2405
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2037..2056)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2247..2268
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2472..2491)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2591..2613
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2651..2669)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2737..2841
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2842..2910,2911..3702)
                     /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(3060..3079)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3873..4461
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4362..4381
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggccttttgc tcacatggct cgacagatct aattctaata cgactcacta tagggaaata
       61 attttgttta actttaagaa ggagatatac cggtatgggc catcaccatc atcaccatca
      121 ccacggaggt ggcactagta tggagctttt cactggcgtt gttcccatcc tggtcgagct
      181 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
      241 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
      301 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
      361 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
      421 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
      481 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
      541 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
      601 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
      661 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
      721 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
      781 ggtcctgctg gagttcgtga ccgccgccgg gatcggaggt ggtggatcgg gcggaggggg
      841 atcaatgcgt atcaacatct ccctgtcgtc gttattcgaa cggttaagta aactgagctc
      901 tcgttcaatt gcgattacct gtggagttgt cctcgcctct gctattgctt ttcccatcat
      961 acgtcgggat tatcagacgt tcctggaggt tggtcctagc tatgccccgc aaaactttcg
     1021 cggctacatt atcgtttgcg tacttagctt gttccgccaa gagcagaaag gcctggcgat
     1081 ctacgaccgg ctcccagaga agcgccgctg gctggcggat ctgccatttc gtgaaggtac
     1141 cagaccgagt attacctcac acattatcca gcgccagcgc actcaacttg tggatcagga
     1201 atttgccaca cgtgaactga ttgacaaggt tattccgagg gtgcaagcac gacataccga
     1261 taaaacattt ttgagcacga gcaagtttga atttcatgcg aaagccattt tcctgttgcc
     1321 atcgattcca atcaacgacc cgctgaacat acctagtcat gatacggtgc gtcgaacgaa
     1381 acgcgaaatt gcccacatgc atgactatca cgattgcacc cttcacctag ctctggcggc
     1441 acaggatgga aaagaagtcc tgaaaaaagg ctggggccag cgtcatccgc tggcggggcc
     1501 gggtgtgcct gggccgccga ctgaatggac tttcttatac gcaccccgta atgaggaaga
     1561 agcgcgcgtg gtagaaatga ttgtggaagc atccatcggc tatatgacca atgatccggc
     1621 aggtaaaatc gtcgagaatg ccaaataaac gcgttaactc gagcaaagcc cgccgatcac
     1681 catcatcacc atcaccacgg aggtggcact agtgaattcc gtacgacgcg ttaactcgag
     1741 caaagcccgc cgaaaggcgg gcttttctgt acaacaacaa ttgcattcat tttatgtttc
     1801 aggttcaggg ggagatgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     1861 aaatcgataa ggatccgggc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     1921 aacagttgcg cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc
     1981 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     2041 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     2101 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     2161 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     2221 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     2281 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     2341 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     2401 aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
     2461 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     2521 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     2581 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     2641 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     2701 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     2761 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     2821 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     2881 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     2941 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     3001 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     3061 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     3121 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     3181 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     3241 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     3301 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     3361 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     3421 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     3481 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     3541 ggagccggtg agcgtgggtc gcgcggtatc attgcagcac tggggccaga tggtaagccc
     3601 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     3661 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     3721 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     3781 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     3841 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3901 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3961 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4021 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4081 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4141 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4201 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4261 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4321 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4381 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4441 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     4501 tgct
//
