LOCUS       sequence-478041-        5080 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing mRuby2 with the CAG promoter and WPRE 3' UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5080)
  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 5080)
  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: "Nucleic 
            Acids Research, Volume 53, Issue 11, 24 June 2025, gkaf528"
COMMENT     Sequence Label: pKG2077_pGEEC518_pShip-CAG-mRuby2-WPRE
FEATURES             Location/Qualifiers
     source          1..5080
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(112..131)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             123..932
                     /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(190..209)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1022..1610
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1511..1530
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      1775..1861
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1953..1980
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     1988..2011
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     enhancer        2110..2413
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3634..3656
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3718..3737
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      3763..3772
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3769..4479
                     /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"
     misc_feature    join(4493..5080,1)
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4546..4566)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4964..4975)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 ccgctcgctg actgcaggtg cccgtgtctc aaaatctctg atgttacatt gcacaagata
       61 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
      121 ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg
      181 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
      241 gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
      301 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
      361 cgaccatcaa gcattttatc cgtactcctg gtgatgcatg gttactcacc actgcgatcc
      421 ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
      481 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
      541 acagcgatcg cgtatttcgt ctagctcagg cgcaatcacg aatgaataac ggtttggttg
      601 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
      661 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
      721 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
      781 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
      841 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
      901 agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacatt
      961 attcagattg ggccccgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     1021 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     1081 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     1141 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     1201 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     1261 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     1321 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     1381 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     1441 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     1501 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     1561 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     1621 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     1681 cgttatcccc tgattctgtg gataaccgta ttaccgctag catggatctc ggggacgtct
     1741 aactactaag cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgga
     1801 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa
     1861 tccgccggga gcggatttga acgttgtgaa gcaacggccc ggagggtggc gggcaggacg
     1921 cccgccataa actgccaggc atcaaactaa gcagaaggcc atcctgacgg atggcctttt
     1981 tgcgtttcta caaactcttc ctgttagtta gttacttaag ctcgggccca cctgcagtct
     2041 acttactact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     2101 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     2161 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     2221 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     2281 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     2341 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     2401 cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc
     2461 ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc
     2521 gggggggggg ggggggcgcg cgccgggggg gggggggggg gggggggggg gggggggggg
     2581 gggggggggg gggggggggg ggggccaggg gggggggggg gccccggagt ttttcttttt
     2641 taacgggggg gggggggggg gggggcttaa aaaaaagagg gcggcgggcg cggggggggc
     2701 ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc
     2761 tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct
     2821 gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt gaaagccttg
     2881 aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg tgcgtgtgtg
     2941 tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag cgctgcgggc
     3001 gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg
     3061 ccccgcggtg cggggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg
     3121 gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca accccccctg cacccccctc
     3181 cccgagttgc tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg
     3241 ggctcgccgt gccgggcggg gggtggcggc aggtgggggt gccgggcggg gcggggccgc
     3301 ctcgggccgg ggagggctcg ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc
     3361 gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac
     3421 ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc accccctcta
     3481 gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg gagggccttc
     3541 gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt ccgcgggggg
     3601 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     3661 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     3721 acgtgctggt tattgtgctg tctcatcatt ttggcaaaaa tggccaccat ggtgtctaag
     3781 ggcgaagagc tgatcaagga aaatatgcgt atgaaggtgg tcatggaagg ttcggtcaac
     3841 ggccaccaat tcaaatgcac aggtgaagga gaaggcaatc cgtacatggg aactcaaacc
     3901 atgaggatca aagtcatcga gggaggaccc ctgccatttg cctttgacat tcttgccacg
     3961 tcgttcatgt atggcagccg tacttttatc aagtacccga aaggcattcc tgatttcttt
     4021 aaacagtcct ttcctgaggg ttttacttgg gaaagagtta cgagatacga agatggtgga
     4081 gtcgtcaccg tcatgcagga caccagcctt gaggatggct gtctcgttta ccacgtccaa
     4141 gtcagagggg taaactttcc ctccaatggt cccgtgatgc agaagaagac caagggttgg
     4201 gagcctaata cagagatgat gtatccagca gatggtggtc tgaggggata cactcatatg
     4261 gcactgaaag ttgatggtgg tggccatctg tcttgctctt tcgtaacaac ttacaggtca
     4321 aaaaagaccg tcgggaacat caagatgccc ggtatccatg ccgttgatca ccgcctggaa
     4381 aggttagagg aaagtgacaa tgaaatgttc gtagtacaac gcgaacacgc agttgccaag
     4441 ttcgccgggc ttggtggtgg gatggacgag ctgtacaagt aacaacgccg ataatcaacc
     4501 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     4561 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     4621 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     4681 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     4741 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     4801 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     4861 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     4921 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     4981 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     5041 ccctcagacg agtcggatct ccctttgggc cgcctccccg
//
