LOCUS       sequence-389877-        6173 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of iRFP713(C15S/V256C) and EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6173)
  TITLE     Plasmids for KOBACHI mice project
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6173)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pCAGGS-iRFP713(C15S/V256C)-P2A-mCherry
FEATURES             Location/Qualifiers
     source          1..6173
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        9..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..665
                     /label=chicken beta-actin promoter
     intron          666..1686
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1606..1628
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1694..1713
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2731..2787
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2788..3492
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTLGMDELY"
     primer_bind     complement(2933..2951)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3201..3220
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3511..3530)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3576..3631
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3630..3649)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3992..4008)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3992..4008)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4005..4027)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4016..4032
                     /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(4040..4046,4047..4064,4065..4070))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4085..4106
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4252..4269)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4423..5011)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4503..4522)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5182..5973,5974..6042))
                     /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     5805..5824
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6043..6147)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgatcga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
       61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      361 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
      421 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
      481 tgcagcgatg ggggcggggg gggggggggs gcgcgccagg cggggcgggg cggggcgagg
      541 ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa
      601 agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc
      661 gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc
      721 gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt
      781 ctcctccggg ctgtaattag cgcttggttt aatgacggct tgtttctttt ctgtggctgc
      841 gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg
      901 cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg
      961 agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg
     1021 ccgggggcgg tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg
     1081 gtgtgtgcgt gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc
     1141 tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg
     1201 ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg
     1261 gggcggggcc gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg
     1321 ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga
     1381 gggcgcaggg acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc
     1441 gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg
     1501 gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct
     1561 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
     1621 gtgtgaccgg cggctctagc tagagcctct gctaaccatg ttcatgcctt cttctttttc
     1681 ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattc
     1741 gtcgacacca tgctcgacct cgagatggcc gaaggatccg tggcaagaca gcccgatctg
     1801 ctgacatctg acgacgaacc aatccacatt ccaggagcta tccagcctca tggactgctg
     1861 ctggctctgg ccgctgacat gactatcgtg gcaggctctg ataacctgcc agagctgacc
     1921 gggctggcta tcggagcact gattggccgg tccgcagccg acgtgttcga ttctgagact
     1981 cacaatagac tgaccatcgc cctggctgaa cctggggctg cagtgggagc cccaattacc
     2041 gtcggcttca caatgcgaaa ggacgctggc tttatcgggt cctggcaccg ccatgatcag
     2101 ctgatttttc tggagctgga accacctcag agagacgtgg cagagccaca ggctttcttt
     2161 cggagaacaa acagcgccat caggcgcctg caggccgctg agactctgga atccgcttgc
     2221 gcagccgctg cacaggaagt gaggaagatc accgggttcg atcgcgtcat gatctacaga
     2281 ttcgcctcag actttagcgg agaagtgatc gcagaagatc gctgtgccga ggtcgaaagt
     2341 aaactgggcc tgcattaccc agcctcaacc gtgcctgcac aggcacgacg gctgtataca
     2401 atcaaccccg tgcggatcat tcccgacatt aattacagac cagtgcccgt cacacctgac
     2461 ctgaacccag tgactgggag gcccatcgat ctgagcttcg ccattctgcg cagcgtgtcc
     2521 ccttgtcacc tggaattcat gaggaatatc ggaatgcatg gcaccatgtc tatcagtatt
     2581 ctgagaggcg aaaggctgtg ggggctgatc gtgtgccacc atcggacacc ttactatgtg
     2641 gacctggacg gcaggcaggc ttgtgaactg gtggctcagg tgctggcttg gcagattggc
     2701 gtgatggaag agggcggccg cggaagcgga gcgacgaatt ttagtctact gaaacaagcg
     2761 ggagacgtgg aggaaaaccc tggacctatg gtgagcaagg gcgaggagga taacatggcc
     2821 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
     2881 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     2941 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
     3001 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
     3061 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
     3121 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     3181 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     3241 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
     3301 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
     3361 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
     3421 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac tctcggcatg
     3481 gacgagctgt acggccgctg aatcgacgga tctttttccc tctgccaaaa attatgggga
     3541 catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc
     3601 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
     3661 tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc
     3721 tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt
     3781 ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt
     3841 tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac tagccagatt
     3901 tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct
     3961 cgacctgcag cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4021 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4081 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4141 gaaacctgtc gtgccagcgg atcagtggac aagctgtact gatccgctgc attaatgaat
     4201 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4261 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4321 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4381 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4441 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4501 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4561 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4621 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4681 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4741 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4801 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4861 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4921 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4981 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5041 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5101 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5161 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5221 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5281 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5341 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5401 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5461 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5521 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5581 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5641 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5701 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5761 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5821 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5881 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5941 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6001 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6061 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6121 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctg
//
