LOCUS       sequence-374562-        4184 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of H3H via a T7 promoter in TXTL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4184)
  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 4184)
  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-H3H
FEATURES             Location/Qualifiers
     source          1..4184
                     /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             1385..1408
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     rep_origin      1630..2085
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1717..1736)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1927..1948
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2152..2171)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2271..2293
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2331..2349)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2417..2521
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2522..2590,2591..3382)
                     /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(2740..2759)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3553..4141
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4042..4061
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggccttttgc tcacatggct cgacagatct aattctaata cgactcacta tagggaaata
       61 attttgttta actttaagaa ggagatatac cggtatgact agtatggcat cctttgaaaa
      121 ttcactcagt gtcttaatag ttggggctgg cttaggtggt ttggccgctg ctatagcact
      181 gcgaaggcaa ggacatgtag tcaagatata cgatagttct tcattcaagg ccgaattggg
      241 agcaggatta gctgtaccac ccaatactct ccgttccctg caacaactgg gctgtaatac
      301 agaaaacctg aacggagtag acaacctctg ctttacggcc atgggatacg acgggagtgt
      361 aggtatgatg aataatatga ccgactaccg tgaggcatac ggcacttcct ggataatggt
      421 acatcgagtg gatctccaca acgagctgat gcgagtcgcc ttggatccgg gcgggttagg
      481 gcctcctgcc actcttcacc ttaaccatcg tgttacgttc tgcgatgtgg atgcctgtac
      541 agtcacgttt acgaacggga cgacacagtc tgctgacttg attgtcggag ctgatggtat
      601 taggtctacg attcgaagat tcgttctgga ggaggacgtg actgttccag ctagcggaat
      661 tgtcggcttt cgatggttgg tccaggccga tgctctcgac ccatatcccg aactggattg
      721 gattgtcaaa aaaccaccct tgggagccag gttgatttct actccacaga acccgcagag
      781 tggcgtagga ctggccgata gacgaacgat cattatttac gcatgcagag gtgggaccat
      841 ggttaatgta ttagccgtac acgacgacga aagagatcaa aacactgctg attggtccgt
      901 tcccgcatca aaggatgatt tattcagagt gtttcacgat taccatccaa gattccgacg
      961 actcttagag ttagcacagg acattaattt atggcagatg agggtggtcc cggtcttgaa
     1021 aaaatgggtg aacaagaggg tatgcttgct gggtgatgcc gcccatgctt cactgcccac
     1081 acttggacaa ggcttcggga tggggttgga ggatgcagtc gctttgggca cgttgttgcc
     1141 taagggaacg accgcatcac agatcgagac aaggctggcc gtttacgaac agctgcgaaa
     1201 ggatcgagct gagtttgtcg ccgcagagag ctatgaggag caatatgtcc cagagatgag
     1261 agggctgtac cttaggtcca aagaattgag agacagagtg atgggttatg atattaaggt
     1321 agagtcagag aaggttttgg aaacactgtt gcgatcatcc aactcagcta cgcgtggagg
     1381 tggccatcac catcatcacc atcaccacgg ataactcgag caaagcccgc cgaaaggcgg
     1441 gcttttctgt acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg
     1501 gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatccgggc
     1561 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     1621 ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     1681 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     1741 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     1801 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     1861 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     1921 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     1981 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     2041 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt
     2101 tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg
     2161 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     2221 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     2281 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     2341 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     2401 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     2461 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     2521 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     2581 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     2641 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     2701 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     2761 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     2821 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     2881 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     2941 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     3001 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     3061 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     3121 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     3181 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     3241 gcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     3301 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     3361 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     3421 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     3481 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     3541 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3601 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3661 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3721 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3781 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3841 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3901 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3961 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4021 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4081 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4141 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgct
//
