LOCUS       sequence-252089-        7760 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of the NIR biosensor for detecting JNK activity
            .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7760)
  AUTHORS   Oliinyk OS, Shemetov AA, Pletnev S, Shcherbakova DM, Verkhusha VV
  TITLE     Smallest near-infrared fluorescent protein evolved from 
            cyanobacteriochrome as versatile tag for spectral multiplexing.
  JOURNAL   Nat Commun. 2019 Jan 17;10(1):279. doi: 10.1038/s41467-018-08050-8.
  PUBMED    30655515
REFERENCE   2  (bases 1 to 7760)
  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 
            Commun."; date: "2019-01-17"; volume: "10(1):279. doi"; pages: " 
            10.1038/s41467-018-08050-8"
COMMENT     Sequence Label: pNIR_JNKbiosensor
FEATURES             Location/Qualifiers
     source          1..7760
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        112..491
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        492..695
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     646..666
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     740..759
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        740..758
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2200..3147
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 720 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP720
                     /note="derived from Rhodopseudomonas palustris RpBphP2"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGSVIAEDRCAEVESKLGLHYPASFIPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPNHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACKRVAERLATQIGVMEE"
     CDS             3160..3186
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     complement(3228..3245)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3234..3458
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3504..3932
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3591..3610)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3801..3822
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3941..3961)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3946..4275
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4126..4261
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4188..4207
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4342..5136
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4396..4415)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5006..5025
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5310..5431
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5347..5366)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5401..5420
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5480..5496)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5480..5496)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5493..5515)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5504..5520
                     /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(5528..5534,5535..5552,5553..5558))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5573..5594
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5711..5728)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5882..6470)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5962..5981)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6641..7432,7433..7501))
                     /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     7264..7283
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7502..7606)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7681..7700)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
       61 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
      121 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      181 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      241 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      301 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      361 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      421 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      481 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      541 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      601 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      661 ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg
      721 cttactggct tatcgaaatt aatacgactc actataggga gacccaagct ggctagcgtt
      781 taaacttaag cttggtaccg agctcggatc cactagtcca gtgtggtgga attcatggca
      841 aacctggaca agatgctgaa taccacagta acagaggtgc ggcagttcct gcaggtggac
      901 agagtgtgcg tgttccagtt tgaggaggat tatagcggag tggtggtggt ggaggccgtg
      961 gacgataggt ggatctccat cctgaagacc caggtgcggg atagatactt catggagaca
     1021 aggggcgagg agtattctca cggccgctac caggccatcg ccgacatcta caccgcaaac
     1081 ctgacagagt gctacaggga tctgctgaca cagtttcagg tgagagcaat cctggccgtg
     1141 cccatcctgc agggcaagaa gctgtggggc ctgttggtgg cacaccagct ggcggcccct
     1201 agacagtggc agacctggga gatcgacttt ctgaagcagc aggccgtggt ggtgggcatc
     1261 gccatccagc agagcctcga gaagttttct caagaacaga tcggcgaaaa cattgtgtgc
     1321 agggtcattt gtaccacggg tcaaattccc atccgagatt tgtcagctga tatttcacaa
     1381 gtgcttaagg aaaaacgatc cataaagaaa gtttggacat ttggtagaaa cccagcctgt
     1441 gactatcatt taggaaacat ttcaagactg tcaaataagc atttccaaat actactagga
     1501 gaagacggta accttttatt gaatgacatt tccactaatg ggacctggtt aaatgggcaa
     1561 aaagtcgaga agaacagcaa tcagttactg tcccaaggtg atgaaataac cgttggtgta
     1621 ggcgtggaat cagatatttt atctctggtc attttcataa acgacaaatt taagcagtgc
     1681 ctggagcaga acaaagttga tcgcggtacc agtgctggtg gtagtgctgg tggtagtgct
     1741 ggtggtagtg ctggtggtag tgctggtggt tccggcagtg ctggtggtag tgctggtggt
     1801 agtaccagtg ctggtggtag tgctggtggt agtgctggtg gtagtgctgg tggtagtgct
     1861 ggtggttccg gcagtgctgg tggtagtgct ggtggtagtg ctggtggtag tgctggtggt
     1921 agtgctggtg gttccggcag tgctggtggt agtgctggtg gtagtaccag tgctggtggt
     1981 agtgctggtg gtagtgctgg tggtagtgct ggtggtagtg ctggtggttc cggcagtgct
     2041 ggtggtagtg ctggtggtag taccagtgct ggtggtagtg ctggtggtag tgctggtggt
     2101 agtgctggtg gtagtgctgg tggttccgga gacagtgtca agacccccga ggatgaaggc
     2161 aacccactgc tagagcagct tgagaagaag ggcggccgca tggcggaagg atccgtcgcc
     2221 aggcagcctg acctcttgac ctgcgacgat gagccgatcc atatccccgg tgccatccaa
     2281 ccgcatggac tgctgctcgc cctcgccgcc gacatgacga tcgttgccgg cagcgacaac
     2341 cttcccgaac tcaccggact ggcgatcggc gccctgatcg gccgctctgc ggccgatgtc
     2401 ttcgactcgg agacgcacaa ccgtctgacg atcgccttgg ccgagcccgg ggcggccgtc
     2461 ggagcaccga tcactgtcgg cttcacgatg cgaaaggacg caggcttcat cggctcctgg
     2521 catcgccatg atcagctcat cttcctggag ctggagcctc cccagcggga cgtcgccgag
     2581 ccgcaggcgt tcttccgccg caccaacagc gccatccgcc gcctgcaggc cgccgaaacc
     2641 ttggaaagcg cctgcgccgc cgcggcgcaa gaggtgcgga agattaccgg cttcgatcgg
     2701 gtgatgatct atcgcttcgc ctccgacttc agcgggtccg tgatcgcaga ggatcggtgc
     2761 gccgaggtcg agtcaaaact aggcctgcac tatcctgcct cattcatccc ggcgcaggcc
     2821 cgtcggctct ataccatcaa cccggtacgg atcattcccg atatcaatta tcggccggtg
     2881 ccggtcaccc cagacctcaa tccggtcacc gggcggccga ttgatcttag cttcgccatc
     2941 ctgcgcagcg tctcgcccaa ccatctggag ttcatgcgca acataggcat gcacggcacg
     3001 atgtcgatct cgattttgcg cggcgagcga ctgtggggat tgatcgtttg ccatcaccga
     3061 acgccgtact acgtcgatct cgatggccgc caagcctgca agagggtcgc cgagaggctg
     3121 gccactcaga tcggcgtgat ggaagagtct agactgcagc tgcctcctct ggaacgcctg
     3181 actctggatt aagtcgacta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     3241 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3301 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3361 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3421 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     3481 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3541 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3601 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3661 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3721 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3781 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3841 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3901 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     3961 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4021 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4081 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4141 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4201 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4261 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     4321 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4381 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4441 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4501 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     4561 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4621 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4681 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4741 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4801 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4861 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4921 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4981 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5041 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5101 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     5161 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     5221 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     5281 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     5341 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5401 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     5461 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     5521 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5581 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5641 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     5701 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5761 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5821 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5881 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5941 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6001 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6061 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6121 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6181 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6241 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6301 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     6361 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6421 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6481 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6541 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6601 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6661 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6721 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6781 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6841 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6901 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6961 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7021 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7081 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7141 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7201 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7261 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7321 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7381 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7441 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7501 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7561 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7621 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     7681 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     7741 ggtcgctgag tagtgcgcga
//
