LOCUS       sequence-366410-        4540 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid can be used to prepare Fluc mRNA by In Vitro 
            Transcription.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4540)
  AUTHORS   Perche F, Clemencon R, Schulze K, Ebensen T, Guzman CA, Pichon C
  TITLE     Neutral Lipopolyplexes for In Vivo Delivery of Conventional and 
            Replicative RNA Vaccine.
  JOURNAL   Mol Ther Nucleic Acids. 2019 Sep 6;17:767-775. doi: 
            10.1016/j.omtn.2019.07.014. Epub 2019 Jul 30.
  PUBMED    31446119
REFERENCE   2  (bases 1 to 4540)
  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: "Mol Ther 
            Nucleic Acids."; date: "2019-09-6"; volume: "17:767-775. doi"; 
            pages: " 10.1016/j.omtn.2019.07.014. Epub 2019 Jul 30"
COMMENT     Sequence Label: pGEM4Z-Fluc
FEATURES             Location/Qualifiers
     source          1..4540
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(242..261)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(421..439)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        507..611
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(612..680,681..1472)
                     /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(830..849)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1643..2231
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2132..2151
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2385..2402
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2519..2540
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2555..2560,2561..2578,2579..2585)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2593..2609
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2598..2620
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2617..2633
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2617..2633
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2652..2671
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2652..2670
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2705..4357
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(2771..2789)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
ORIGIN
        1 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaawaaaa aaaaaaaaaa
       61 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
      121 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaactag tggcgcctga
      181 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
      241 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
      301 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
      361 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
      421 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
      481 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
      541 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
      601 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
      661 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
      721 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
      781 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
      841 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
      901 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
      961 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     1021 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     1081 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     1141 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     1201 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     1261 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     1321 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     1381 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     1441 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     1501 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     1561 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     1621 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     1681 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     1741 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     1801 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     1861 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     1921 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     1981 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     2041 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     2101 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     2161 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     2221 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     2281 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     2341 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     2401 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     2461 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     2521 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     2581 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     2641 acgccaagct ctaatacgac tcactatagg gagacaagct tggcattccg gtactgttgg
     2701 taaaatggaa gacgccaaaa acataaagaa aggcccggcg ccattctatc ctctagagga
     2761 tggaaccgct ggagagcaac tgcataaggc tatgaagaga tacgccctgg ttcctggaac
     2821 aattgctttt acagatgcac atatcgaggt gaacatcacg tacgcggaat acttcgaaat
     2881 gtccgttcgg ttggcagaag ctatgaaacg atatgggctg aatacaaatc acagaatcgt
     2941 cgtatgcagt gaaaactctc ttcaattctt tatgccggtg ttgggcgcgt tatttatcgg
     3001 agttgcagtt gcgcccgcga acgacattta taatgaacgt gaattgctca acagtatgaa
     3061 catttcgcag cctaccgtag tgtttgtttc caaaaagggg ttgcaaaaaa ttttgaacgt
     3121 gcaaaaaaaa ttaccaataa tccagaaaat tattatcatg gattctaaaa cggattacca
     3181 gggatttcag tcgatgtaca cgttcgtcac atctcatcta cctcccggtt ttaatgaata
     3241 cgattttgta ccagagtcct ttgatcgtga caaaacaatt gcactgataa tgaattcctc
     3301 tggatctact gggttaccta agggtgtggc ccttccgcat agaactgcct gcgtcagatt
     3361 ctcgcatgcc agagatccta tttttggcaa tcaaatcatt ccggatactg cgattttaag
     3421 tgttgttcca ttccatcacg gttttggaat gtttactaca ctcggatatt tgatatgtgg
     3481 atttcgagtc gtcttaatgt atagatttga agaagagctg tttttacgat cccttcagga
     3541 ttacaaaatt caaagtgcgt tgctagtacc aaccctattt tcattcttcg ccaaaagcac
     3601 tctgattgac aaatacgatt tatctaattt acacgaaatt gcttctgggg gcgcacctct
     3661 ttcgaaagaa gtcggggaag cggttgcaaa acgcttccat cttccaggga tacgacaagg
     3721 atatgggctc actgagacta catcagctat tctgattaca cccgaggggg atgataaacc
     3781 gggcgcggtc ggtaaagttg ttccattttt tgaagcgaag gttgtggatc tggataccgg
     3841 gaaaacgctg ggcgttaatc agagaggcga attatgtgtc agaggaccta tgattatgtc
     3901 cggttatgta aacaatccgg aagcgaccaa cgccttgatt gacaaggatg gatggctaca
     3961 ttctggagac atagcttact gggacgaaga cgaacacttc ttcatagttg accgcttgaa
     4021 gtctttaatt aaatacaaag gatatcaggt ggcccccgct gaattggaat cgatattgtt
     4081 acaacacccc aacatcttcg acgcgggcgt ggcaggtctt cccgacgatg acgccggtga
     4141 acttcccgcc gccgttgttg ttttggagca cggaaagacg atgacggaaa aagagatcgt
     4201 ggattacgtc gccagtcaag taacaaccgc gaaaaagttg cgcggaggag ttgtgtttgt
     4261 ggacgaagta ccgaaaggtc ttaccggaaa actcgacgca agaaaaatca gagagatcct
     4321 cataaaggcc aagaagggcg gaaagtccaa attgtaaaat gtaactgtat tcagcgatga
     4381 cgaaattctt agctattgta atattatatg caaattgatg aatggtaatt ttgtaattgt
     4441 gggtcactgt actattttaa cgaataataa aatcaggtat aggtaactaa aaaggaattc
     4501 gagctcgaat tcttaattaa aaaaaaaaaa aaaaaaaaaa
//
