LOCUS       sequence-475956-        3406 bp    DNA     circular SYN 10-JUL-2025
DEFINITION  Plasmid expressing mRuby from a UbC promoter lacking an intron, used
            for modRNA production.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3406)
  AUTHORS   Peterman EL, Ploessl DS, Love KS, Sanabria V, Daniels RF, Johnstone 
            CP, Godavarti DR, Kabaria SR, Oakes CG, Pai AA, Galloway KE
  TITLE     High-resolution profiling reveals coupled transcriptional and 
            translational regulation of transgenes.
  JOURNAL   Nucleic Acids Research, Volume 53, Issue 11, 24 June 2025, gkaf528
  PUBMED    39651241
REFERENCE   2  (bases 1 to 3406)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 10, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Research, Volume 53, Issue 11, 24 June 2025, gkaf528"
COMMENT     Sequence Label: pKG3831_pGEEC579_pShip-UbC(no_intron)-mRuby2-bGH
FEATURES             Location/Qualifiers
     source          1..3406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      463..472
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             469..1179
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     polyA_signal    1194..1418
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1529..1548)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1540..2349
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPGDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1607..1626)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2439..3027
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2928..2947
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      3192..3278
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3370..3397
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     join(3405..3406,1..22)
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
ORIGIN
        1 acaaactctt cctgttagtt agttacttaa gctcgggccc acctgcagtc tacttactgg
       61 cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc tcctcacggc gagcgctgcc
      121 acgtcagacg aagggcgcag cgagcgtcct gatccttccg cccggacgct caggacagcg
      181 gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga cattttagga
      241 cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac aggcgaggaa
      301 aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga acgccgatga
      361 ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg atttgggtcg
      421 cggttcttgt ttgtggatcg ctgtgatcgt cacttgacaa tggccaccat ggtgtctaag
      481 ggcgaagagc tgatcaagga aaatatgcgt atgaaggtgg tcatggaagg ttcggtcaac
      541 ggccaccaat tcaaatgcac aggtgaagga gaaggcaatc cgtacatggg aactcaaacc
      601 atgaggatca aagtcatcga gggaggaccc ctgccatttg cctttgacat tcttgccacg
      661 tcgttcatgt atggcagccg tacttttatc aagtacccga aaggcattcc tgatttcttt
      721 aaacagtcct ttcctgaggg ttttacttgg gaaagagtta cgagatacga agatggtgga
      781 gtcgtcaccg tcatgcagga caccagcctt gaggatggct gtctcgttta ccacgtccaa
      841 gtcagagggg taaactttcc ctccaatggt cccgtgatgc agaagaagac caagggttgg
      901 gagcctaata cagagatgat gtatccagca gatggtggtc tgaggggata cactcatatg
      961 gcactgaaag ttgatggtgg tggccatctg tcttgctctt tcgtaacaac ttacaggtca
     1021 aaaaagaccg tcgggaacat caagatgccc ggtatccatg ccgttgatca ccgcctggaa
     1081 aggttagagg aaagtgacaa tgaaatgttc gtagtacaac gcgaacacgc agttgccaag
     1141 ttcgccgggc ttggtggtgg gatggacgag ctgtacaagt aacaacgttg cacctgtgcc
     1201 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     1261 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     1321 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     1381 caatagcagg catgctgggg atgcggtggg ctctatggcg ctcgctgact gcaggtgccc
     1441 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
     1501 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     1561 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     1621 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
     1681 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     1741 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     1801 actcctggtg atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta
     1861 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     1921 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtcta
     1981 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     2041 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     2101 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     2161 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     2221 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     2281 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     2341 tttttctaat cagaattggt taattggttg taacattatt cagattgggc cccgttccac
     2401 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2461 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2521 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2581 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2641 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2701 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2761 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2821 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2881 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2941 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     3001 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     3061 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     3121 aaccgtatta ccgctagcat ggatctcggg gacgtctaac tactaagcga gagtagggaa
     3181 ctgccaggca tcaaataaaa cgaaaggctc agtcggaaga ctgggccttt cgttttatct
     3241 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg
     3301 ttgtgaagca acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc
     3361 aaactaagca gaaggccatc ctgacggatg gcctttttgc gtttct
//
