LOCUS       sequence-177136-        4881 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  pCAG promoter driving mRuby2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4881)
  AUTHORS   Weinberg BH, Pham NT, Caraballo LD, Lozanoski T, Engel A, Bhatia S, 
            Wong WW
  TITLE     Large-scale design of robust genetic circuits with multiple inputs 
            and outputs for mammalian cells.
  JOURNAL   Nat Biotechnol. 2017 Mar 27. doi: 10.1038/nbt.3805.
  PUBMED    28346402
REFERENCE   2  (bases 1 to 4881)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2017 Mar 27. doi: 10.1038/nbt.3805."
COMMENT     Sequence Label: pCAG-mRuby2
FEATURES             Location/Qualifiers
     source          1..4881
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     466..488
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     550..569
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      626..635
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             632..1342
                     /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"
     primer_bind     complement(1431..1450)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1496..1551
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1550..1569)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1912..1928)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1912..1928)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1925..1947)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1936..1952
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1960..1966,1967..1984,1985..1990))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2005..2026
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      2111..2245
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2265..2399
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2302..2321)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2356..2375
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2467..2484)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2638..3226)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2718..2737)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3397..4188,4189..4257))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4020..4039
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4258..4362)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        4393..4772
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg
       61 ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg
      121 gggcggggcc gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg
      181 ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga
      241 gggcgcaggg acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc
      301 gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg
      361 gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct
      421 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
      481 gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac
      541 agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattgatta
      601 attcgagcga acgcgtaccg gtgccgccac catggtgtct aagggcgaag agctgatcaa
      661 ggaaaatatg cgtatgaagg tggtcatgga aggttcggtc aacggccacc aattcaaatg
      721 cacaggtgaa ggagaaggca atccgtacat gggaactcaa accatgagga tcaaagtcat
      781 cgagggagga cccctgccat ttgcctttga cattcttgcc acgtcgttca tgtatggcag
      841 ccgtactttt atcaagtacc cgaaaggcat tcctgatttc tttaaacagt cctttcctga
      901 gggttttact tgggaaagag ttacgagata cgaagatggt ggagtcgtca ccgtcatgca
      961 ggacaccagc cttgaggatg gctgtctcgt ttaccacgtc caagtcagag gggtaaactt
     1021 tccctccaat ggtcccgtga tgcagaagaa gaccaagggt tgggagccta atacagagat
     1081 gatgtatcca gcagatggtg gtctgagggg atacactcat atggcactga aagttgatgg
     1141 tggtggccat ctgtcttgct ctttcgtaac aacttacagg tcaaaaaaga ccgtcgggaa
     1201 catcaagatg cccggtatcc atgccgttga tcaccgcctg gaaaggttag aggaaagtga
     1261 caatgaaatg ttcgtagtac aacgcgaaca cgcagttgcc aagttcgccg ggcttggtgg
     1321 tgggatggac gagctgtaca agtaagcggc cgcactcctc aggtgcaggc tgcctatcag
     1381 aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc
     1441 tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa
     1501 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
     1561 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca
     1621 acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat
     1681 atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag
     1741 atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta
     1801 catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct
     1861 cttctcttat ggagatccct cgacctgcag cccaagcttg gcgtaatcat ggtcatagct
     1921 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     1981 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     2041 actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct caattagtca
     2101 gcaaccatag tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2161 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     2221 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctaacttg tttattgcag
     2281 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     2341 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc
     2401 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     2461 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2521 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2581 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2641 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2701 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2761 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2821 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2881 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2941 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3001 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3061 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3121 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3181 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3241 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3301 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3361 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3421 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3481 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     3541 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     3601 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     3661 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     3721 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     3781 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     3841 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     3901 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     3961 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4021 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4081 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4141 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4201 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4261 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4321 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4381 gccacctggg tcgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
     4441 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     4501 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     4561 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     4621 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     4681 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     4741 acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttca
     4801 ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat
     4861 tttgtgcagc gatgggggcg g
//
