LOCUS       sequence-369379-        4250 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Endogenous murine RNase inhibitor production for cell-free lysate.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4250)
  AUTHORS   Voyvodic PL, Conejero I, Mesmoudi K, Renard E, Courtet P, Cattoni 
            DI, Bonnet J
  TITLE     Evaluating and mitigating clinical samples matrix effects on TX-TL 
            cell-free performance.
  JOURNAL   Sci Rep. 2022 Aug 12;12(1):13785. doi: 10.1038/s41598-022-17583-4.
  PUBMED    35962056
REFERENCE   2  (bases 1 to 4250)
  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: "2022-08-12"; volume: "12(1):13785. doi"; pages: " 
            10.1038/s41598-022-17583-4"
COMMENT     Sequence Label: pPLV_C1
FEATURES             Location/Qualifiers
     source          1..4250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      148..175
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     276..295
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        276..294
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             326..348
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2065..2082)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2236..2824)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2316..2335)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2995..3786,3787..3855))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3618..3637
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3856..3960)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4028..4046
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cgcctgatgc gtgaacgtga cggacgtaac caccgcgaca tgtgtgtgct gttccgctgg
       61 ctcggatacc cttactctgt tgaaaacgaa tagataggtt aaggaacggt tatttctgcg
      121 tagatctatc ttacacagca tcacactggc tcaccttcgg gtgggccttt ctgcgtttat
      181 atactagaga gagaatataa aaagccagat tattaatccg gcttttttat tatttaggca
      241 actgaaacga ttcggatcct gtattactat tcttataata cgactcacta tagggagacc
      301 acaacggttt ccctctagaa ataattttgt ttaactttaa gaaggagata taccatgtcg
      361 ttagacattc aatgcgagca gctgtcggac gcccgttgga cggaattatt accactgatc
      421 caacaatatg aggtggtacg tttggatgat tgcgggttga ccgaagttcg ctgtaaagat
      481 atctcatctg ccgtgcaggc taacccggcc ttaactgagc ttagcctgcg taccaatgag
      541 ctgggtgacg gcggtgtggg tttggtgctg cagggtctgc aaaacccgac gtgcaaaatc
      601 caaaaactgt cacttcagaa ttgtggactc actgaggcag gttgcggcat tctgcccggt
      661 atgcttcgta gtcttagcac cctccgcgaa ctgcatctga atgataatcc gatgggcgac
      721 gcgggcctga aattgttatg tgaaggtctg caggatccac aatgtcgcct ggagaaactg
      781 caattagagt actgcaacct taccgcgacg tcttgcgaac cgctggcctc agtcttgcgc
      841 gtaaaagcgg atttcaaaga attggtgctg tcgaacaacg atctgcatga accaggggtc
      901 cgtatcttgt gccagggctt aaaggattcc gcctgccagc tcgagtccct caaactggaa
      961 aactgtggta tcaccgcagc taattgtaaa gatctctgcg acgtggtggc atccaaggct
     1021 tcgttacagg aactggatct ctcttctaat aaattgggta atgcaggaat cgcagcctta
     1081 tgccctggcc ttctcttgcc gagttgtaaa ttacgcacct tgtggctgtg ggaatgcgat
     1141 atcacggcgg aaggttgtaa agacctgtgt cgtgtgctgc gggccaagca gtcactgaaa
     1201 gaattatcgc tggcctcgaa cgaactgaaa gatgagggcg cgcgtttatt gtgcgagtca
     1261 ttactggagc caggatgcca gctggaatca ctgtggatta aaacctgcag cttgaccgca
     1321 gcgtcgtgcc cgtacttctg tagtgtgctg accaaatcgc gcagcttgct tgagctgcag
     1381 atgtcttcca atccccttgg tgacgaaggt gtgcaagaac tttgcaaggc tttgagtcag
     1441 ccggacaccg tactccgcga actgtggctg ggggattgtg atgtcaccaa ctctggttgt
     1501 tcatccttgg cgaatgttct gctggcgaac cgtagcctgc gtgagttaga tctgagtaac
     1561 aattgtatgg gtggcccagg tgtgctgcag ctgttagaaa gtctgaagca accgtcttgt
     1621 accctgcagc aactggttct gtatgatatt tattggacaa acgaggtcga agagcaactg
     1681 cgcgcgctgg aagaagagcg cccgagttta cggatcattt cctgataaac tttatctgag
     1741 aatagtcaat cttcggaaat cccaggtggc atgctaaaag tctcgtaaag cgttctatca
     1801 ataacccgtt ggtgcaaagc ccgccgaaag gcgggctttt ctgtccgtct cagaatcggc
     1861 cgtgaacaat aaaatagttt cggtattatt gaccacttcc gagtagaatc gtgcttcagt
     1921 aagagtcgac cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
     1981 ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
     2041 gtgccggcag cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     2101 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2161 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2221 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2281 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2341 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2401 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2461 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2521 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2581 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2641 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2701 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2761 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2821 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2881 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2941 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3001 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3061 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     3121 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3181 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3241 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3301 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3361 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3421 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3481 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3541 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3601 cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3661 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3721 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3781 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     3841 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     3901 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     3961 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     4021 ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa gaattctggc gaatcctctg
     4081 accagccaga aaacgacctt tctgtggtga aaccggatgc tgcaattcag agcgccagca
     4141 agtgggggac agcagaagac ctgaccgccg cagagtggat gtttgacatg gtgaagacta
     4201 tcgcaccatc agccagaaaa ccgaattttg ctgggtgggc taacgatatc
//
