LOCUS       sequence-389068-        5676 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the protein of mRhubarb720 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5676)
  AUTHORS   Zhang H, Papadaki S, Sun X, Wang X, Drobizhev M, Yao L, Rehbock M, 
            Koster RW, Wu L, Namikawa K, Piatkevich KD
  TITLE     Quantitative assessment of near-infrared fluorescent proteins.
  JOURNAL   Nat Methods. 2023 Sep 4. doi: 10.1038/s41592-023-01975-z.
  PUBMED    37666982
REFERENCE   2  (bases 1 to 5676)
  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. 2023 Sep 4. doi: 10.1038/s41592-023-01975-z."
COMMENT     Sequence Label: pCAG-mRhubarb720
FEATURES             Location/Qualifiers
     source          1..5676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        281..558
                     /label=chicken beta-actin promoter
     primer_bind     837..859
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     921..940
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      979..988
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             985..1935
                     /codon_start=1
                     /product="phytochrome-based near-infrared fluorescent 
                     protein, originally termed iRFP (Filonov et al., 2011)"
                     /label=iRFP713
                     /note="name was changed to iRFP713 (Shcherbakova and 
                     Verkhusha, 2013)"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRHTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGEVIAEDRCAEVESKLGQHYPASDIPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPVHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLARAIGVMEE"
     misc_feature    1960..2548
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2013..2033)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2431..2442)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2591..2712
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2602..2621)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2656..2675
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3240..3828)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3320..3339)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3999..4790,4791..4859))
                     /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     4622..4641
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4860..4964)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4990..5445)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5127..5148)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5339..5358
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tatctacgta gccatgctct aggaagagta ccattgacgt caataatgac gtatgttccc
       61 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      121 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      181 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      241 tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc
      301 tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt
      361 taattatttt gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc aggcggggcg
      421 gggcggggcg aggggcgggg cggggcgagg cggagaggtg cggcggcagc caatcagagc
      481 ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag
      541 cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc
      601 gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg
      661 cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg gcttgtttct
      721 tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg cggggggagc
      781 ggctcggggc tgtccgcggg gggacggctg ccttcggggg ggacggggca gggcggggtt
      841 cggcttctgg cgtgtgaccg gcggctctag agcctctgct aaccatgttc atgccttctt
      901 ctttttccta cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc attttggcaa
      961 agaattggat ccggtaccgc caccatggct gaaggatccg tcgccaggca gcctgacctc
     1021 ttgacctgcg acgatgagcc gatccatatc cccggtgcca tccaaccgca tggactgctg
     1081 ctcgccctcg ccgccgacat gacgatcgtt gccggcagcg acaaccttcc cgaactcacc
     1141 ggactggcga tcggcgccct gatcggccgc tctgcggccg atgtcttcga ctcggagacg
     1201 cacaaccgtc tgacgatcgc cttggccgag cccggggcgg ccgtcggagc accgatcact
     1261 gtcggcttca cgatgcgaaa ggacgcaggc ttcatcggct cctggcatcg ccatgatcag
     1321 ctcatcttcc tcgagctcga gcctccccag cgggacgtcg ccgagccgca ggcgttcttc
     1381 cgccacacca acagcgccat ccgccgcctg caggccgccg aaaccttgga gagcgcctgc
     1441 gccgccgcgg cgcaagaggt gcggaagatt accggcttcg atcgggtgat gatctatcgc
     1501 ttcgcctccg acttcagcgg cgaagtgatc gcagaggatc ggtgcgccga ggtcgagtca
     1561 aaactaggcc agcactatcc tgcctcagat attccggcgc aggcccgtcg gctctatacc
     1621 atcaacccgg tacggatcat tcccgatatc aattatcggc cggtgccggt caccccagac
     1681 ctcaatccgg tcaccgggcg gccgattgat cttagcttcg ccatcctgcg cagcgtctcg
     1741 cccgtccatc tggaattcat gcgcaacata ggcatgcacg gcacgatgtc gatctcgatt
     1801 ttgcgcggcg agcgactgtg gggattgatc gtttgccatc accgaacgcc gtactacgtc
     1861 gatctcgatg gccgccaagc ctgcgagcta gtcgcccagg ttctggcccg ggcgatcggc
     1921 gtgatggaag agtaagaatt cgatatcaag cttatcgata atcaacctct ggattacaaa
     1981 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     2041 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     2101 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     2161 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     2221 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     2281 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     2341 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
     2401 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     2461 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     2521 cggatctccc tttgggccgc ctccccgcat cgataccgtc gacccgggcg gccgcttcga
     2581 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     2641 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     2701 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg
     2761 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatctt
     2821 cctagagcat ggctacgtag ataagtagca tggcgggtta atcattaact acaaggaacc
     2881 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgsscg
     2941 rscwwagstc gsccgrcgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     3001 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     3061 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     3121 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3181 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3241 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3301 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3361 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3421 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3481 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3541 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3601 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3661 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3721 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3781 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3841 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3901 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3961 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4021 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4081 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4141 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4201 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4261 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     4321 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4381 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     4441 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4501 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4561 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     4621 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4681 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4741 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4801 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4861 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4921 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4981 gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     5041 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     5101 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     5161 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     5221 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     5281 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     5341 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     5401 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca
     5461 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgc
     5521 gcgctcgctc gctcactgag gccgggcgac ctttggtcgg gcgtcgggcg acctttggtc
     5581 gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg
     5641 gttccttgta gttaatgatt aacccgccat gctact
//
