LOCUS       sequence-387461-        5639 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the protein of iRFP670 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5639)
  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 5639)
  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: pAAV-CAG-iRFP670
FEATURES             Location/Qualifiers
     source          1..5639
                     /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..1920
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 670 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP670
                     /note="derived from Rhodopseudomonas palustris RpBphP6"
                     /translation="MARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAG
                     AFFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLH
                     RHDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVM
                     LYRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSR
                     IVPEHDASGAALDLSFAHLRSISPCHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYE
                     PRAVPMAQRVAAEMFADFLSLHFTAAHHQR"
     misc_feature    1945..2533
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1998..2018)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2416..2427)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2576..2697
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2587..2606)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2641..2660
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3203..3791)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3283..3302)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3962..4753,4754..4822))
                     /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     4585..4604
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4823..4927)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4953..5408)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5090..5111)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5302..5321
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5480..5609
                     /label=AAV2 ITR
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 ggsgcgcgcc 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 caccatggcg cgtaaggtcg atctcacctc ctgcgatcgc
     1021 gagccgatcc acatccccgg cagcattcag ccgtgcggct gcctgctagc ctgcgacgcg
     1081 caggcggtgc ggatcacgcg cattacggaa aatgccggcg cgttctttgg acgcgaaact
     1141 ccgcgggtcg gtgagctact cgccgattac ttcggcgaga ccgaagccca tgcgctgcgc
     1201 aacgcactgg cgcagtcctc cgatccaaag cgaccggcgc tgatcttcgg ttggcgcgac
     1261 ggcctgaccg gccgcacctt cgacatctca ctgcatcgcc atgacggtac atcgatcatc
     1321 gagttcgagc ctgcggcggc cgaacaggcc gacaatccgc tgcggctgac gcggcagatc
     1381 atcgcgcgca ccaaagaact gaagtcgctc gaagagatgg ccgcacgggt gccgcgctat
     1441 ctgcaggcga tgctcggcta tcaccgcgtg atgttgtacc gcttcgcgga cgacggctcc
     1501 gggatggtga tcggcgaggc gaagcgcagc gacctcgaga gctttctcgg tcagcacttt
     1561 ccggcgtcgc tggtcccgca gcaggcgcgg ctactgtact tgaagaacgc gatccgcgtg
     1621 gtctcggatt cgcgcggcat cagcagccgg atcgtgcccg agcacgacgc ctccggcgcc
     1681 gcgctcgatc tgtcgttcgc gcacctgcgc agcatctcgc cctgccatct cgaatttctg
     1741 cggaacatgg gcgtcagcgc ctcgatgtcg ctgtcgatca tcattgacgg cacgctatgg
     1801 ggattgatca tctgtcatca ttacgagccg cgtgccgtgc cgatggcgca gcgcgtcgcg
     1861 gccgaaatgt tcgccgactt cttatcgctg cacttcaccg ccgcccacca ccaacgctaa
     1921 gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     1981 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     2041 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     2101 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     2161 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     2221 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     2281 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     2341 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     2401 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     2461 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     2521 gccgcctccc cgcatcgata ccgtcgaccc gggcggccgc ttcgagcaga catgataaga
     2581 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     2641 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     2701 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa
     2761 agcaagtaaa acctctacaa atgtggtaaa atcgataagg atcttcctag agcatggcta
     2821 cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt
     2881 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gcccgggctt tgcccgggcg
     2941 gcctcagtga gcgagcgagc gcgcagctgc attaatgaat cggccaacgc gcggggagag
     3001 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3061 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3121 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3181 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3241 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3301 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3361 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3421 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3481 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3541 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3601 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3661 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3721 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3781 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3841 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3901 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3961 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4021 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4081 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4141 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4201 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4261 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4321 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4381 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4441 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4501 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4561 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4621 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     4681 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     4741 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4801 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     4861 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     4921 ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa
     4981 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     5041 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     5101 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     5161 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     5221 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     5281 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     5341 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     5401 gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     5461 ctcttcgcta ttacgccagc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc
     5521 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
     5581 agtggccaac tccatcacta ggggttcctt gtagttaatg attaacccgc catgctact
//
