LOCUS       sequence-117252-        4111 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IVT vector for Fluc mRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4111)
  AUTHORS   Andries O, Cafferty SM, De Smedt SC, Weiss R, Sanders NN, Kitada T
  TITLE     N-methylpseudouridine-incorporated mRNA outperforms 
            pseudouridine-incorporated mRNA by providing enhanced protein 
            expression and reduced immunogenicity in mammalian cell lines and 
            mice.
  JOURNAL   J Control Release. 2015 Sep 2. pii: S0168-3659(15)30094-8. doi: 
            10.1016/j.jconrel.2015.08.051.
  PUBMED    26342664
REFERENCE   2  (bases 1 to 4111)
  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: "J Control 
            Release. 2015 Sep 2. pii: S0168-3659(15)30094-8. doi: 
            10.1016/j.jconrel.2015.08.051."
COMMENT     Sequence Label: pTK305
FEATURES             Location/Qualifiers
     source          1..4111
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        19..37
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    74..98
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      99..108
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             105..1757
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     protein_bind    complement(1758..1782)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     rep_origin      complement(2357..2945)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2437..2456)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3116..3907,3908..3976))
                     /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     3739..3758
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3977..4081)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tagggataac agggtaatta atacgactca ctataggatg gactacgaca tagtctagtc
       61 cgccaagtct agaacaagtt tgtacaaaaa agcaggctgc caccatggaa gatgccaaaa
      121 acattaagaa gggcccagcg ccattctacc cactcgaaga cgggaccgcc ggcgagcagc
      181 tgcacaaagc catgaagcgc tacgccctgg tgcccggcac catcgccttt accgacgcac
      241 atatcgaggt ggacattacc tacgccgagt acttcgagat gagcgttcgg ctggcagaag
      301 ctatgaagcg ctatgggctg aatacaaacc atcggatcgt ggtgtgcagc gagaatagct
      361 tgcagttctt catgcccgtg ttgggtgccc tgttcatcgg tgtggctgtg gccccagcta
      421 acgacatcta caacgagcgc gagctgctga acagcatggg catcagccag cccaccgtcg
      481 tattcgtgag caagaaaggg ctgcaaaaga tcctcaacgt gcaaaagaag ctaccgatca
      541 tacaaaagat catcatcatg gatagcaaga ccgactacca gggcttccaa agcatgtaca
      601 ccttcgtgac ttcccatttg ccacccggct tcaacgagta cgacttcgtg cccgagagct
      661 tcgaccggga caaaaccatc gccctgatca tgaacagtag tggcagtacc ggattgccca
      721 agggcgtagc cctaccgcac cgcaccgctt gtgtccgatt cagtcatgcc cgcgacccca
      781 tcttcggcaa ccagatcatc cccgacaccg ctatcctcag cgtggtgcca tttcaccacg
      841 gcttcggcat gttcaccacg ctgggctact tgatctgcgg ctttcgggtc gtgctcatgt
      901 accgcttcga ggaggagcta ttcttgcgca gcttgcaaga ctataagatt caatctgccc
      961 tgctggtgcc cacactattt agcttcttcg ctaagagcac tctcatcgac aagtacgacc
     1021 taagcaactt gcacgagatc gccagcggcg gggcgccgct cagcaaggag gtaggtgagg
     1081 ccgtggccaa acgcttccac ctaccaggca tccgccaggg ctacggcctg acagaaacaa
     1141 ccagcgccat tctgatcacc cccgaagggg acgacaagcc tggcgcagta ggcaaggtgg
     1201 tgcccttctt cgaggctaag gtggtggact tggacaccgg taagacactg ggtgtgaacc
     1261 agcgcggcga gctgtgcgtc cgtggcccca tgatcatgag cggctacgtt aacaaccccg
     1321 aggctacaaa cgctctcatc gacaaggacg gctggctgca cagcggcgac atcgcctact
     1381 gggacgagga cgagcacttc ttcatcgtgg accggctgaa gagcctgatc aaatacaagg
     1441 gctaccaggt agccccagcc gaactggaga gcatcctgct gcaacacccc aacatcttcg
     1501 acgccggggt cgccggcctg cccgacgacg atgccggcga gctgcccgcc gcagtcgtcg
     1561 tgctggaaca cggtaaaacc atgaccgaga aggagatcgt ggactatgtg gccagccagg
     1621 ttacaaccgc caagaagctg cgcggtggtg ttgtgttcgt ggacgaggtg cctaaaggac
     1681 tgaccggcaa gttggacgcc cgcaagatcc gcgagattct cattaaggcc aagaagggcg
     1741 gcaagatcgc cgtgtaaacc cagctttctt gtacaaagtg gtggcgcgcc atacagcagc
     1801 aattggcaag ctgcttacat agaactcgcg gcgattggca tgccgcctta aaatttttat
     1861 tttatttttc ttttcttttc cgaatcggat tttgttttta atatttccgc gcctatgtta
     1921 cgtgcaaagg tgattgtcac cccccgaaag accatattgt gacacaccct cagtatcacg
     1981 cccaaacatt tacagccgcg gtgtcaaaaa ccgcgtggac gtggttaaca tccctgctgg
     2041 gaggatcagc cgtaattatt ataattggct tggtgctggc tactattgtg gccatgtacg
     2101 tgctgaccaa ccagaaacat aattgaatac agcagcaatt ggcaagctgc ttacatagaa
     2161 ctcgcggcga ttggcatgcc gccttaaaat ttttatttta tttttctttt cttttccgaa
     2221 tcggattttg tttttaatat ttcaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     2281 aatagggata acagggtaat tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2341 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2401 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2461 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2521 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2581 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2641 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2701 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2761 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     2821 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     2881 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     2941 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3001 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3061 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3121 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3181 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3241 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3301 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3361 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     3421 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     3481 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     3541 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     3601 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     3661 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     3721 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     3781 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     3841 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     3901 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     3961 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4021 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4081 gcacatttcc ccgaaaagtg ccacctgacg t
//
