LOCUS       sequence-475940-        2513 bp    DNA     circular SYN 10-JUL-2025
DEFINITION  BsaI Golden Gate assembly plasmid encoding the mRuby2 CDS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2513)
  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 2513)
  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: pKG0586_pPV2-mRuby2
FEATURES             Location/Qualifiers
     source          1..2513
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        41..145
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(146..214,215..1006)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGYRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(364..383)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1177..1765
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1666..1685
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     regulatory      1790..1799
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1796..2506
                     /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"
ORIGIN
        1 agagacctag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
       61 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
      121 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
      181 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
      241 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
      301 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
      361 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
      421 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
      481 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
      541 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
      601 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
      661 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
      721 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
      781 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
      841 atctggagcc ggtgagcgtg ggtatcgcgg tatcattgca gcactggggc cagatggtaa
      901 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
      961 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     1021 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     1081 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     1141 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     1201 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     1261 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     1321 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     1381 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     1441 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     1501 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     1561 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     1621 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     1681 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     1741 gtcagggggg cggagcctat ggaaaaacgc cagcaacggg tctcaaatgg ccaccatggt
     1801 gtctaagggc gaagagctga tcaaggaaaa tatgcgtatg aaggtggtca tggaaggttc
     1861 ggtcaacggc caccaattca aatgcacagg tgaaggagaa ggcaatccgt acatgggaac
     1921 tcaaaccatg aggatcaaag tcatcgaggg aggacccctg ccatttgcct ttgacattct
     1981 tgccacgtcg ttcatgtatg gcagccgtac ttttatcaag tacccgaaag gcattcctga
     2041 tttctttaaa cagtcctttc ctgagggttt tacttgggaa agagttacga gatacgaaga
     2101 tggtggagtc gtcaccgtca tgcaggacac cagccttgag gatggctgtc tcgtttacca
     2161 cgtccaagtc agaggggtaa actttccctc caatggtccc gtgatgcaga agaagaccaa
     2221 gggttgggag cctaatacag agatgatgta tccagcagat ggtggtctga ggggatacac
     2281 tcatatggca ctgaaagttg atggtggtgg ccatctgtct tgctctttcg taacaactta
     2341 caggtcaaaa aagaccgtcg ggaacatcaa gatgcccggt atccatgccg ttgatcaccg
     2401 cctggaaagg ttagaggaaa gtgacaatga aatgttcgta gtacaacgcg aacacgcagt
     2461 tgccaagttc gccgggcttg gtggtgggat ggacgagctg tacaagtaac aac
//
