LOCUS       sequence-475976-        3598 bp    DNA     circular SYN 10-JUL-2025
DEFINITION  Plasmid expressing mRuby2 from the CMV canonical promoter with the 
            hPGK 5' UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3598)
  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 3598)
  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: 
            pKG3780_pGEEC587_pShip-CMV(promoter)-hPGK(UTR)-mRuby2-bGH
FEATURES             Location/Qualifiers
     source          1..3598
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     7..30
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     enhancer        67..370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        371..574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     525..545
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      663..672
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             669..1379
                     /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    1394..1618
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1729..1748)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1740..2549
                     /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(1807..1826)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2639..3227
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3128..3147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      3392..3478
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3570..3597
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gcgtttctac aaactcttcc tgttagttag ttacttaagc tcgggcccac ctgcagtcta
       61 cttactcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      541 gcgtgtacgg tgggaggtct atataagcag agctgagctc gtttagtgaa ccgtcctccg
      601 gagcgcacgt cggcagtcgg ctccctcgtt gaccgaatca ccgacctctc tccccagaaa
      661 tggccaccat ggtgtctaag ggcgaagagc tgatcaagga aaatatgcgt atgaaggtgg
      721 tcatggaagg ttcggtcaac ggccaccaat tcaaatgcac aggtgaagga gaaggcaatc
      781 cgtacatggg aactcaaacc atgaggatca aagtcatcga gggaggaccc ctgccatttg
      841 cctttgacat tcttgccacg tcgttcatgt atggcagccg tacttttatc aagtacccga
      901 aaggcattcc tgatttcttt aaacagtcct ttcctgaggg ttttacttgg gaaagagtta
      961 cgagatacga agatggtgga gtcgtcaccg tcatgcagga caccagcctt gaggatggct
     1021 gtctcgttta ccacgtccaa gtcagagggg taaactttcc ctccaatggt cccgtgatgc
     1081 agaagaagac caagggttgg gagcctaata cagagatgat gtatccagca gatggtggtc
     1141 tgaggggata cactcatatg gcactgaaag ttgatggtgg tggccatctg tcttgctctt
     1201 tcgtaacaac ttacaggtca aaaaagaccg tcgggaacat caagatgccc ggtatccatg
     1261 ccgttgatca ccgcctggaa aggttagagg aaagtgacaa tgaaatgttc gtagtacaac
     1321 gcgaacacgc agttgccaag ttcgccgggc ttggtggtgg gatggacgag ctgtacaagt
     1381 aacaacgttg cacctgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     1441 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     1501 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     1561 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggcg
     1621 ctcgctgact gcaggtgccc gtgtctcaaa atctctgatg ttacattgca caagataaaa
     1681 atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta
     1741 tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt
     1801 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct
     1861 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     1921 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
     1981 ccatcaagca ttttatccgt actcctggtg atgcatggtt actcaccact gcgatccccg
     2041 gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     2101 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
     2161 gcgatcgcgt atttcgtcta gctcaggcgc aatcacgaat gaataacggt ttggttgatg
     2221 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     2281 ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     2341 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
     2401 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
     2461 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
     2521 ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg taacattatt
     2581 cagattgggc cccgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2641 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2701 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2761 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     2821 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2881 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2941 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3001 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3061 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3121 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3181 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3241 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     3301 tatcccctga ttctgtggat aaccgtatta ccgctagcat ggatctcggg gacgtctaac
     3361 tactaagcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcggaaga
     3421 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     3481 gccgggagcg gatttgaacg ttgtgaagca acggcccgga gggtggcggg caggacgccc
     3541 gccataaact gccaggcatc aaactaagca gaaggccatc ctgacggatg gccttttt
//
