LOCUS       sequence-476959-        3222 bp    DNA     circular SYN 24-JUN-2025
DEFINITION  Plasmid expressing mRuby2 from the EFS canonical promoter with the 
            CMV 5' UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3222)
  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   bioRxiv [Preprint]. 2024 Dec 3:2024.11.26.625483. doi: 
            10.1101/2024.11.26.625483.
  PUBMED    39651241
REFERENCE   2  (bases 1 to 3222)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 24, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2024 Dec 3:2024.11.26.625483. doi: 
            10.1101/2024.11.26.625483."
COMMENT     Sequence Label: 
            pKG3777_pGEEC584_pShip-EFS(promoter)-CMV(UTR)-mRuby2-bGH
FEATURES             Location/Qualifiers
     source          1..3222
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      2..29
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     37..60
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     regulatory      317..326
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             323..1033
                     /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
                     MALKVDGSGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     polyA_signal    1048..1272
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1383..1402)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1394..2203
                     /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(1461..1480)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2293..2881
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2782..2801
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      3046..3132
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cagaaggcca tcctgacgga tggccttttt gcgtttctac aaactcttcc tgttagttag
       61 ttacttaagc tcgggcccac ctgcagtcta cttactgggc agagcgcaca tcgcccacag
      121 tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg
      181 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
      241 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgagatcgc ctggagacgc
      301 catccacgct gtaatggcca ccatggtgtc taagggcgaa gagctgatca aggaaaatat
      361 gcgtatgaag gtggtcatgg aaggttcggt caacggccac caattcaaat gcacaggtga
      421 aggagaaggc aatccgtaca tgggaactca aaccatgagg atcaaagtca tcgagggagg
      481 acccctgcca tttgcctttg acattcttgc cacgtcgttc atgtatggca gccgtacttt
      541 tatcaagtac ccgaaaggca ttcctgattt ctttaaacag tcctttcctg agggttttac
      601 ttgggaaaga gttacgagat acgaagatgg tggagtcgtc accgtcatgc aggacaccag
      661 ccttgaggat ggctgtctcg tttaccacgt ccaagtcaga ggggtaaact ttccctccaa
      721 tggtcccgtg atgcagaaga agaccaaggg ttgggagcct aatacagaga tgatgtatcc
      781 agcagatggt ggtctgaggg gatacactca tatggcactg aaagttgatg gtagtggcca
      841 tctgtcttgc tctttcgtaa caacttacag gtcaaaaaag accgtcggga acatcaagat
      901 gcccggtatc catgccgttg atcaccgcct ggaaaggtta gaggaaagtg acaatgaaat
      961 gttcgtagta caacgcgaac acgcagttgc caagttcgcc gggcttggtg gtgggatgga
     1021 cgagctgtac aagtaacaac gttgcacctg tgccttctag ttgccagcca tctgttgttt
     1081 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     1141 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     1201 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     1261 tgggctctat ggcgctcgct gactgcaggt gcccgtgtct caaaatctct gatgttacat
     1321 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa
     1381 tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa
     1441 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     1501 gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     1561 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     1621 tatgcctctt ccgaccatca agcattttat ccgtactcct ggtgatgcat ggttactcac
     1681 cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     1741 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     1801 ttgtcctttt aacagcgatc gcgtatttcg tctagctcag gcgcaatcac gaatgaataa
     1861 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     1921 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     1981 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     2041 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     2101 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     2161 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
     2221 gttgtaacat tattcagatt gggccccgtt ccactgagcg tcagaccccg tagaaaagat
     2281 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2341 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2401 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     2461 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2521 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2581 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     2641 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     2701 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     2761 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     2821 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     2881 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     2941 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcta gcatggatct
     3001 cggggacgtc taactactaa gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag
     3061 gctcagtcgg aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
     3121 agtaggacaa atccgccggg agcggatttg aacgttgtga agcaacggcc cggagggtgg
     3181 cgggcaggac gcccgccata aactgccagg catcaaacta ag
//
