LOCUS       sequence-475957-        3561 bp    DNA     circular SYN 10-JUL-2025
DEFINITION  Plasmid expressing mRuby2 from the CMV canonical promoter with the 
            EFS 5' UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3561)
  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 3561)
  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: 
            pKG3779_pGEEC586_pShip-CMV(promoter)-EFS(UTR)-mRuby2-bGH
FEATURES             Location/Qualifiers
     source          1..3561
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     16..39
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     enhancer        76..379
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        380..583
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     534..554
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      635..644
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             641..1351
                     /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    1366..1590
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1701..1720)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1712..2521
                     /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(1779..1798)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2611..3199
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3100..3119
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      3364..3450
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(3542..3561,1..8)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggcccacc
       61 tgcagtctac ttactcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      121 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      181 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      241 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      301 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      361 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      421 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      481 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      541 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctgagctcg tttagtgaac
      601 cgtccaacgg gtttgccgcc agaacacagt aatggccacc atggtgtcta agggcgaaga
      661 gctgatcaag gaaaatatgc gtatgaaggt ggtcatggaa ggttcggtca acggccacca
      721 attcaaatgc acaggtgaag gagaaggcaa tccgtacatg ggaactcaaa ccatgaggat
      781 caaagtcatc gagggaggac ccctgccatt tgcctttgac attcttgcca cgtcgttcat
      841 gtatggcagc cgtactttta tcaagtaccc gaaaggcatt cctgatttct ttaaacagtc
      901 ctttcctgag ggttttactt gggaaagagt tacgagatac gaagatggtg gagtcgtcac
      961 cgtcatgcag gacaccagcc ttgaggatgg ctgtctcgtt taccacgtcc aagtcagagg
     1021 ggtaaacttt ccctccaatg gtcccgtgat gcagaagaag accaagggtt gggagcctaa
     1081 tacagagatg atgtatccag cagatggtgg tctgagggga tacactcata tggcactgaa
     1141 agttgatggt ggtggccatc tgtcttgctc tttcgtaaca acttacaggt caaaaaagac
     1201 cgtcgggaac atcaagatgc ccggtatcca tgccgttgat caccgcctgg aaaggttaga
     1261 ggaaagtgac aatgaaatgt tcgtagtaca acgcgaacac gcagttgcca agttcgccgg
     1321 gcttggtggt gggatggacg agctgtacaa gtaacaacgt tgcacctgtg ccttctagtt
     1381 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     1441 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     1501 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     1561 ggcatgctgg ggatgcggtg ggctctatgg cgctcgctga ctgcaggtgc ccgtgtctca
     1621 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
     1681 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag
     1741 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     1801 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt
     1861 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     1921 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctgg
     1981 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga
     2041 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     2101 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tagctcaggc
     2161 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     2221 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     2281 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     2341 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     2401 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     2461 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     2521 atcagaattg gttaattggt tgtaacatta ttcagattgg gccccgttcc actgagcgtc
     2581 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2641 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2701 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     2761 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2821 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2881 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2941 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3001 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3061 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3121 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3181 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3241 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3301 taccgctagc atggatctcg gggacgtcta actactaagc gagagtaggg aactgccagg
     3361 catcaaataa aacgaaaggc tcagtcggaa gactgggcct ttcgttttat ctgttgtttg
     3421 tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgtgaag
     3481 caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaactaag
     3541 cagaaggcca tcctgacgga t
//
