LOCUS       sequence-475974-        3463 bp    DNA     circular SYN 10-JUL-2025
DEFINITION  Plasmid expressing mRuby2 from a SynZiFTR promoter, Induced with 
            pKG2309.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3463)
  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 3463)
  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: 
            pKG3660_pGEEC570_pShip-ZF10.BSx8-ybTATA-spacer-mRuby2-bGH
FEATURES             Location/Qualifiers
     source          1..3463
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      82..670
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     571..590
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      835..921
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1013..1040
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     1048..1071
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     regulatory      1569..1578
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1575..2285
                     /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    2300..2524
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2635..2654)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2646..3455
                     /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(2713..2732)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 ggttaattgg ttgtaacatt attcagattg ggccccgttc cactgagcgt cagaccccgt
       61 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
      121 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
      181 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
      241 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
      301 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
      361 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
      421 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
      481 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
      541 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
      601 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
      661 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
      721 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgctag
      781 catggatctc ggggacgtct aactactaag cgagagtagg gaactgccag gcatcaaata
      841 aaacgaaagg ctcagtcgga agactgggcc tttcgtttta tctgttgttt gtcggtgaac
      901 gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgtgaa gcaacggccc
      961 ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaactaa gcagaaggcc
     1021 atcctgacgg atggcctttt tgcgtttcta caaactcttc ctgttagtta gttacttaag
     1081 ctcgggccca cctgcagtct acttactgac gattgagtaa cactgtatct agtctaccta
     1141 cagggctcgg cgtagccgat gtcgcgcgga tagcgtcaac ctcttcggcg tagccgatgt
     1201 cgcgcaatca gttctcccga ggtcggcgta gccgatgtcg cgcagagccg tgcttccaat
     1261 acggcgtagc cgatgtcgcg ccgggtttcg taacaatcgc atgaggattc gcaacgcctt
     1321 cggcgtagcc gatgtcgcgc tcccgtctca gtaaaggtcg gcgtagccga tgtcgcgcaa
     1381 tcggactgcc ttcgtacggc gtagccgatg tcgcgcgtat cagtcgcctc ggaacggcgt
     1441 agccgatgtc gcgcattcgt aagaggctca ctctccctta cacggagtgg ataggatcct
     1501 ctagagggta tataatgggg gccactagtc tactaccaga gctcatcgct agcgctaccg
     1561 gatcaatggc caccatggtg tctaagggcg aagagctgat caaggaaaat atgcgtatga
     1621 aggtggtcat ggaaggttcg gtcaacggcc accaattcaa atgcacaggt gaaggagaag
     1681 gcaatccgta catgggaact caaaccatga ggatcaaagt catcgaggga ggacccctgc
     1741 catttgcctt tgacattctt gccacgtcgt tcatgtatgg cagccgtact tttatcaagt
     1801 acccgaaagg cattcctgat ttctttaaac agtcctttcc tgagggtttt acttgggaaa
     1861 gagttacgag atacgaagat ggtggagtcg tcaccgtcat gcaggacacc agccttgagg
     1921 atggctgtct cgtttaccac gtccaagtca gaggggtaaa ctttccctcc aatggtcccg
     1981 tgatgcagaa gaagaccaag ggttgggagc ctaatacaga gatgatgtat ccagcagatg
     2041 gtggtctgag gggatacact catatggcac tgaaagttga tggtggtggc catctgtctt
     2101 gctctttcgt aacaacttac aggtcaaaaa agaccgtcgg gaacatcaag atgcccggta
     2161 tccatgccgt tgatcaccgc ctggaaaggt tagaggaaag tgacaatgaa atgttcgtag
     2221 tacaacgcga acacgcagtt gccaagttcg ccgggcttgg tggtgggatg gacgagctgt
     2281 acaagtaaca acgttgcacc tgtgccttct agttgccagc catctgttgt ttgcccctcc
     2341 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     2401 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     2461 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     2521 atggcgctcg ctgactgcag gtgcccgtgt ctcaaaatct ctgatgttac attgcacaag
     2581 ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
     2641 gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg
     2701 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
     2761 atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
     2821 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
     2881 ttccgaccat caagcatttt atccgtactc ctggtgatgc atggttactc accactgcga
     2941 tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
     3001 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
     3061 ttaacagcga tcgcgtattt cgtctagctc aggcgcaatc acgaatgaat aacggtttgg
     3121 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
     3181 aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
     3241 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
     3301 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
     3361 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
     3421 tgcagtttca tttgatgctc gatgagtttt tctaatcaga att
//
