LOCUS       sequence-475975-        3256 bp    DNA     circular SYN 10-JUL-2025
DEFINITION  Plasmid expressing mRuby2 from the EFS canonical promoter with the 
            hPGK 5' UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3256)
  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 3256)
  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: 
            pKG3778_pGEEC585_pShip-EFS(promoter)-hPGK(UTR)-mRuby2-bGH
FEATURES             Location/Qualifiers
     source          1..3256
                     /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      351..360
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             357..1067
                     /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    1082..1306
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1417..1436)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1428..2237
                     /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(1495..1514)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2327..2915
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2816..2835
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      3080..3166
                     /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 tcgctccgga gcgcacgtcg
      301 gcagtcggct ccctcgttga ccgaatcacc gacctctctc cccagaaatg gccaccatgg
      361 tgtctaaggg cgaagagctg atcaaggaaa atatgcgtat gaaggtggtc atggaaggtt
      421 cggtcaacgg ccaccaattc aaatgcacag gtgaaggaga aggcaatccg tacatgggaa
      481 ctcaaaccat gaggatcaaa gtcatcgagg gaggacccct gccatttgcc tttgacattc
      541 ttgccacgtc gttcatgtat ggcagccgta cttttatcaa gtacccgaaa ggcattcctg
      601 atttctttaa acagtccttt cctgagggtt ttacttggga aagagttacg agatacgaag
      661 atggtggagt cgtcaccgtc atgcaggaca ccagccttga ggatggctgt ctcgtttacc
      721 acgtccaagt cagaggggta aactttccct ccaatggtcc cgtgatgcag aagaagacca
      781 agggttggga gcctaataca gagatgatgt atccagcaga tggtggtctg aggggataca
      841 ctcatatggc actgaaagtt gatggtggtg gccatctgtc ttgctctttc gtaacaactt
      901 acaggtcaaa aaagaccgtc gggaacatca agatgcccgg tatccatgcc gttgatcacc
      961 gcctggaaag gttagaggaa agtgacaatg aaatgttcgt agtacaacgc gaacacgcag
     1021 ttgccaagtt cgccgggctt ggtggtggga tggacgagct gtacaagtaa caacgttgca
     1081 cctgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     1141 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     1201 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     1261 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcgct cgctgactgc
     1321 aggtgcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat
     1381 gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc
     1441 aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata
     1501 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc
     1561 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag
     1621 atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt
     1681 ttatccgtac tcctggtgat gcatggttac tcaccactgc gatccccgga aaaacagcat
     1741 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt
     1801 tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat
     1861 ttcgtctagc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg
     1921 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc
     1981 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg
     2041 acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc
     2101 aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc
     2161 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc
     2221 tcgatgagtt tttctaatca gaattggtta attggttgta acattattca gattgggccc
     2281 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     2341 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2401 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     2461 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     2521 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     2581 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     2641 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     2701 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     2761 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     2821 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     2881 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     2941 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     3001 ctgtggataa ccgtattacc gctagcatgg atctcgggga cgtctaacta ctaagcgaga
     3061 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcggaagact gggcctttcg
     3121 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga
     3181 tttgaacgtt gtgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc
     3241 caggcatcaa actaag
//
