LOCUS       sequence-169621-        4767 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses CyRFP1 fluorescent protein in an empty backbone, under CAG
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4767)
  AUTHORS   Laviv T, Kim BB, Chu J, Lam AJ, Lin MZ, Yasuda R
  TITLE     Simultaneous dual-color fluorescence lifetime imaging with novel 
            red-shifted fluorescent proteins.
  JOURNAL   Nat Methods. 2016 Oct 31. doi: 10.1038/nmeth.4046.
  PUBMED    27798609
REFERENCE   2  (bases 1 to 4767)
  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 
            Methods. 2016 Oct 31. doi: 10.1038/nmeth.4046."
COMMENT     Sequence Label: pCAG-CyRFP1
FEATURES             Location/Qualifiers
     source          1..4767
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     335..357
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     423..442
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      486..495
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             492..1193
                     /codon_start=1
                     /product="cyan-excitable red fluorescent protein (Laviv et 
                     al., 2016)"
                     /label=CyRFP1
                     /translation="MVSKGEELIKENMRSKLYLEGSVNGHQFKCTHEGEGKPYEGKQTN
                     RIKVVEGGPLPFAFDILATMFMYGSKVFIKYPADLPDYFKQSFPEGFTWERVMVFEDGG
                     VLTATQDTSLQDGELIYNVKLRGVNFPANGPVMQKKTLGWEPSTETMYPADGGLEGRCD
                     KALKLVGGGHLHVNFKTTYKSKKPVKMPGVHYVDRRLERIKEADNETYVEQYEHAVARY
                     SNLGGGMDELYK"
     misc_feature    1194..1259
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     complement(1289..1308)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1354..1409
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1408..1427)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1770..1786)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1770..1786)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1783..1805)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1794..1810
                     /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(1818..1824,1825..1842,1843..1848))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1863..1884
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1943..2138
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1989..2124
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2144..2278
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2181..2200)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2235..2254
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2346..2363)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2517..3105)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2597..2616)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3276..4067,4068..4136))
                     /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     3899..3918
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4137..4241)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        4276..4655
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cctcgggccg gggagggctc gggggagggg cgcggcggcc cccggagcgc cggcggctgt
       61 cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga
      121 cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct
      181 agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
      241 cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg
      301 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
      361 ggctctagct agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct
      421 gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaattcg ctagcgctac
      481 cggtcgccac catggtgagc aagggcgagg agctgatcaa ggagaacatg agaagcaagc
      541 tgtacctgga aggcagcgtg aacggccacc agttcaagtg cacccacgaa ggggagggca
      601 agccctacga gggcaagcag accaacagga tcaaggtggt ggagggaggc cccctgccgt
      661 tcgcattcga catcctggcc accatgttta tgtacgggag caaggtgttc atcaagtacc
      721 ccgccgacct ccccgattat tttaagcagt ccttccctga gggcttcaca tgggagagag
      781 tcatggtgtt cgaagacggg ggcgtgctga ccgccaccca ggacaccagc ctccaggacg
      841 gcgagctcat ctacaacgtc aagctcagag gggtgaactt cccagccaac ggccccgtga
      901 tgcagaagaa aacactgggc tgggagccca gcaccgagac catgtacccc gctgacggcg
      961 gcctggaagg cagatgcgac aaggccctga agctcgtggg cgggggccac ctgcacgtca
     1021 acttcaagac cacatacaag tccaagaaac ccgtgaagat gcccggcgtc cactacgtgg
     1081 accgcagact ggaaagaatc aaggaggccg acaacgagac ctacgtcgag cagtacgagc
     1141 acgctgtggc cagatactcc aacctgggcg gaggcatgga cgagctgtac aagtccggac
     1201 tcagatctcg agctcaagct tcgaattctg cagtcgacgg taccgcgggc ccgggatcct
     1261 aagcggccgc gactctagag tcgacggatc tttttccctc tgccaaaaat tatggggaca
     1321 tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa
     1381 tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt
     1441 taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg
     1501 ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc
     1561 attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg
     1621 tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt
     1681 tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg
     1741 acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     1801 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     1861 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     1921 aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc ccgcccctaa
     1981 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     2041 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     2101 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc
     2161 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     2221 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccg
     2281 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2341 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2401 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2461 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2521 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2581 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2641 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2701 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2761 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2821 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     2881 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     2941 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3001 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3061 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3121 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3181 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3241 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3301 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3361 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3421 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3481 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3541 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     3601 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     3661 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     3721 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     3781 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     3841 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     3901 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     3961 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4021 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4081 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4141 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4201 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4261 ccacctgggt cgatcgacat tgattattga ctagttatta atagtaatca attacggggt
     4321 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     4381 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     4441 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     4501 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     4561 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     4621 agtacatcta cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct
     4681 tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt attttttaat
     4741 tattttgtgc agcgatgggg gcggggg
//
