LOCUS       sequence-362894-        7466 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cytosolic iRFP702 expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7466)
  AUTHORS   Yang JM, Chi WY, Liang J, Takayanagi S, Iglesias PA, Huang CH
  TITLE     Deciphering cell signaling networks with massively multiplexed 
            biosensor barcoding.
  JOURNAL   Cell. 2021 Dec 9;184(25):6193-6206.e14. doi: 
            10.1016/j.cell.2021.11.005. Epub 2021 Nov 26.
  PUBMED    34838160
REFERENCE   2  (bases 1 to 7466)
  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"; doi: 
            "10.1016/j.cell.2021.11.005"; journalName: "Cell"; date: "2021-12-9-
            9"; volume: "184"; issue: "25"; pages: "6193-6206.e14"
COMMENT     Sequence Label: pVITRO1-iRFP702-NES
FEATURES             Location/Qualifiers
     source          1..7466
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    302..423
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(313..332)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     367..386
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      611..1199
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1100..1119
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1279..1582
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1697..3002
                     /label=rEF-1-alpha promoter
                     /note="strong constitutive promoter for rat elongation 
                     factor EF-1-alpha"
     misc_feature    2039..2970
                     /label=rEF-1-alpha intron
                     /note="intron upstream of the start codon of rat 
                     EF-1-alpha"
     regulatory      3009..3018
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3015..3947
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 702 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP702
                     /note="derived from Rhodopseudomonas palustris RpBphP6"
                     /translation="MARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAG
                     AFFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLH
                     RHDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVM
                     LYRFADDGSGKVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSR
                     IVPEHDASGAALDLSFAHLRSISPIHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYE
                     PRAVPMAQRVAAEMFADFLSLHFTAAHHQR"
     misc_feature    4008..4452
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        4485..4531
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        4523..4541
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4551..5576
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    5588..6160
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
     enhancer        6212..6403
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6235..6255)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6417..7466,1..257)
                     /label=mEF-1-alpha promoter
                     /note="strong constitutive promoter for mouse elongation 
                     factor EF-1-alpha"
     intron          join(6744..7466,1..225)
                     /label=mEF-1-alpha intron
                     /note="intron upstream of the start codon of mouse 
                     EF-1-alpha"
ORIGIN
        1 ggtcctatcg gccgcaatag tcacctcggg cttttgagca cggctagtcg cggcgggggg
       61 aggggatgta atggcgttgg agtttgttca catttggtgg gtggagacta gtcaggccag
      121 cctggcgctg gaagtcattt ttggaatttg tccccttgag ttttgagcgg agctaattct
      181 cgggcttctt agcggttcaa aggtatcttt taaacccttt tttaggtgtt gtgaaaacca
      241 ccgctaattc aaagcaaccg gtgatatcaa agatcttgta cagctagctg gccagacatg
      301 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
      361 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
      421 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
      481 ttttaaagca agtaaaacct ctacaaatgt ggtatggaaa tgttaattaa ctagccatga
      541 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
      601 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
      661 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
      721 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
      781 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
      841 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
      901 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
      961 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     1021 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     1081 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     1141 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     1201 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     1261 tcttaattaa cctgcaggcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     1321 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     1381 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     1441 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     1501 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     1561 attagtcatc gctattacca tgatgatgcg gttttggcag tacatcaatg ggcgtggata
     1621 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     1681 ttgactagtg gagccgagag taattcatac aaaaggaggg atcgccttcg caaggggaga
     1741 gcccagggac cgtccctaaa ttctcacaga cccaaatccc tgtagccgcc ccacgacagc
     1801 gcgaggagca tgcgcccagg gctgagcgcg ggtagatcag agcacacaag ctcacagtcc
     1861 ccggcggtgg ggggaggggc gcgctgagcg ggggccaggg agctggcgcg gggcaaactg
     1921 ggaaagtggt gtcgtgtgct ggctccgccc tcttcccgag ggtgggggag aacggtatat
     1981 aagtgcggta gtcgccttgg acgttctttt tcgcaacggg tttgccgtca gaacgcaggt
     2041 gagtggcggg tgtggcttcc gcgggccccg gagctggagc cctgctctga gcgggccggg
     2101 ctgatatgcg agtgtcgtcc gcagggttta gctgtgagca ttcccacttc gagtggcggg
     2161 cggtgcgggg gtgagagtgc gaggcctagc ggcaaccccg tagcctcgcc tcgtgtccgg
     2221 cttgaggcct agcgtggtgt ccgccgccgc gtgccactcc ggccgcacta tgcgtttttt
     2281 gtccttgctg ccctcgattg ccttccagca gcatgggcta acaaagggag ggtgtggggc
     2341 tcactcttaa ggagcccatg aagcttacgt tggataggaa tggaagggca ggaggggcga
     2401 ctggggcccg cccgccttcg gagcacatgt ccgacgccac ctggatgggg cgaggcctgt
     2461 ggctttccga agcaatcggg cgtgagttta gcctacctgg gccatgtggc cctagcactg
     2521 ggcacggtct ggcctggcgg tgccgcgttc ccttgcctcc caacaagggt gaggccgtcc
     2581 cgcccggcac cagttgcttg cgcggaaaga tggccgctcc cggggccctg ttgcaaggag
     2641 ctcaaaatgg aggacgcggc agcccggtgg agcgggcggg tgagtcaccc acacaaagga
     2701 agagggcctt gcccctcgcc ggccgctgct tcctgtgacc ccgtggtcta tcggccgcat
     2761 agtcacctcg ggcttctctt gagcaccgct cgtcgcggcg gggggagggg atctaatggc
     2821 gttggagttt gttcacattt ggtgggtgga gactagtcag gccagcctgg cgctggaagt
     2881 cattcttgga atttgcccct ttgagtttgg agcgaggcta attctcaagc ctcttagcgg
     2941 ttcaaaggta ttttctaaac ccgtttccag gtgttgtgaa agccaccgct aattcaaagc
     3001 aatccggagc caccatggcg cgtaaggtcg atctcacctc ctgcgatcgc gagccgatcc
     3061 acatccccgg cagcattcag ccgtgcggct gcctgctagc ctgcgacgcg caggcggtgc
     3121 ggatcacgcg cattacggaa aatgccggcg cgttctttgg acgcgaaact ccgcgggtcg
     3181 gtgagctact cgccgattac ttcggcgaga ccgaagccca tgcgctgcgc aacgcactgg
     3241 cgcagtcctc cgatccaaag cgaccggcgc tgatcttcgg ttggcgcgac ggcctgaccg
     3301 gccgcacctt cgacatctcg ctgcatcgcc atgacggtac atcgatcatc gagttcgagc
     3361 ctgcggcggc cgaacaggcc gacaatccgc tgcggctgac gcggcagatc atcgcgcgca
     3421 ccaaagaact gaagtcgctc gaagagatgg ccgcacgggt gccgcgctat ctgcaggcga
     3481 tgctcggcta tcaccgcgtg atgttgtacc gcttcgcgga cgacggctcc ggcaaagtga
     3541 tcggcgaggc gaagcgcagc gacctcgaga gctttctcgg tcagcacttt ccggcgtcgc
     3601 tggtcccgca gcaggcgcgg ctactgtact tgaagaacgc gatccgcgtg gtctcggatt
     3661 cgcgcggcat cagcagccgg atcgtgcccg agcacgacgc ctccggcgcc gcgcttgatc
     3721 tgtcgttcgc gcacctgcgc agcatctcgc ctatccatct cgaatttctg cggaacatgg
     3781 gcgtcagcgc ctcgatgtcg ctgtcgatca tcattgacgg cacgctatgg ggattgatca
     3841 tctgtcatca ttacgagccg cgtgccgtgc cgatggcgca gcgcgtcgcg gccgaaatgt
     3901 tcgccgactt cttatcgctg cacttcaccg ccgcccacca ccaacgcgga agctccctcc
     3961 agaagaaact ggaggaactg gaactgtgag gatcccgtac gcctaggagc aggtttcccc
     4021 aatgacacaa aacgtgcaac ttgaaactcc gcctggtctt tccaggtcta gaggggtaac
     4081 actttgtact gcgtttggct ccacgctcga tccactggcg agtgttagta acagcactgt
     4141 tgcttcgtag cggagcatga cggccgtggg aactcctcct tggtaacaag gacccacggg
     4201 gccaaaagcc acgcccacac gggcccgtca tgtgtgcaac cccagcacgg cgactttact
     4261 gcgaaaccca ctttaaagtg acattgaaac tggtacccac acactggtga caggctaagg
     4321 atgcccttca ggtaccccga ggtaacacgc gacactcggg atctgagaag gggactgggg
     4381 cttctataaa agcgctcggt ttaaaaagct tctatgcctg aataggtgac cggaggtcgg
     4441 cacctttcct ttgcaattac tgaccctatg aatacactga ctgtttgaca attaatcatc
     4501 ggcatagtat atcggcatag tataatacga ctcactatag gagggccacc atgaagaaac
     4561 ctgaactgac agcaacttct gttgagaagt ttctcattga aaaatttgat tctgtttctg
     4621 atctcatgca gctgtctgaa ggtgaagaaa gcagagcctt ttcttttgat gttggaggaa
     4681 gaggttatgt tctgagggtc aattcttgtg ctgatggttt ttacaaagac agatatgttt
     4741 acagacactt tgcctctgct gctctgccaa ttccagaagt tctggacatt ggagaatttt
     4801 ctgaatctct cacctactgc atcagcagaa gagcacaagg agtcactctc caggatctcc
     4861 ctgaaactga gctgccagct gttctgcaac ctgttgctga agcaatggat gccattgcag
     4921 cagctgatct gagccaaacc tctggatttg gtccttttgg tccccaaggc attggtcagt
     4981 acaccacttg gagggatttc atttgtgcca ttgctgatcc tcatgtctat cactggcaga
     5041 ctgtgatgga tgacacagtt tctgcttctg ttgctcaggc actggatgaa ctcatgctgt
     5101 gggcagaaga ttgtcctgaa gtcagacacc tggtccatgc tgattttgga agcaacaatg
     5161 ttctgacaga caatggcaga atcactgcag tcattgactg gtctgaagcc atgtttggag
     5221 attctcaata tgaggttgcc aacatttttt tttggagacc ttggctggct tgcatggaac
     5281 aacaaacaag atattttgaa agaagacacc cagaactggc tggttccccc agactgagag
     5341 cctacatgct cagaattggc ctggaccaac tgtatcaatc tctggttgat ggaaactttg
     5401 atgatgctgc ttgggcacaa ggaagatgtg atgccattgt gaggtctggt gctggaactg
     5461 ttggaagaac tcaaattgca agaaggtctg ctgctgtttg gactgatgga tgtgttgaag
     5521 ttctggctga ctctggaaac aggagaccct ccacaagacc cagagccaag gaatgaatat
     5581 tagctagatt atccctaata cctgccaccc cactcttaat cagtggtgga agaacggtct
     5641 cagaactgtt tgtttcaatt ggccatttaa gtttagtagt aaaagactgg ttaatgataa
     5701 caatgcatcg taaaaccttc agaaggaaag gagaatgttt tgtggaccac tttggttttc
     5761 ttttttgcgt gtggcagttt taagttatta gtttttaaaa tcagtacttt ttaatggaaa
     5821 caacttgacc aaaaatttgt cacagaattt tgagacccat taaaaaagtt aaatgagaaa
     5881 cctgtgtgtt cctttggtca acaccgagac atttaggtga aagacatcta attctggttt
     5941 tacgaatctg gaaacttctt gaaaatgtaa ttcttgagtt aacacttctg ggtggagaat
     6001 agggttgttt tccccccaca taattggaag gggaaggaat atcatttaaa gctatgggag
     6061 ggttgctttg attacaacac tggagagaaa tgcagcatgt tgctgattgc ctgtcactaa
     6121 aacaggccaa aaactgagtc cttgggttgc atagaaagct gcctgcaggg cctgaaataa
     6181 cctctgaaag aggaacttgg ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg
     6241 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     6301 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     6361 gtatgcaaag catgcatctc aattagtcag caaccatagt cccactagtg gagccgagag
     6421 taattcatac aaaaggaggg atcgccttcg caaggggaga gcccagggac cgtccctaaa
     6481 ttctcacaga cccaaatccc tgtagccgcc ccacgacagc gcgaggagca tgcgctcagg
     6541 gctgagcgcg gggagagcag agcacacaag ctcatagacc ctggtcgtgg gggggaggac
     6601 cggggagctg gcgcggggca aactgggaaa gcggtgtcgt gtgctggctc cgccctcttc
     6661 ccgagggtgg gggagaacgg tatataagtg cggcagtcgc cttggacgtt ctttttcgca
     6721 acgggtttgc cgtcagaacg caggtgaggg gcgggtgtgg cttccgcggg ccgccgagct
     6781 ggaggtcctg ctccgagcgg gccgggcccc gctgtcgtcg gcggggatta gctgcgagca
     6841 ttcccgcttc gagttgcggg cggcgcggga ggcagagtgc gaggcctagc ggcaaccccg
     6901 tagcctcgcc tcgtgtccgg cttgaggcct agcgtggtgt ccgcgccgcc gccgcgtgct
     6961 actccggccg cactctggtc tttttttttt ttgttgttgt tgccctgctg ccttcgattg
     7021 ccgttcagca ataggggcta acaaagggag ggtgcggggc ttgctcgccc ggagcccgga
     7081 gaggtcatgg ttggggagga atggagggac aggagtggcg gctggggccc gcccgccttc
     7141 ggagcacatg tccgacgcca cctggatggg gcgaggcctg gggtttttcc cgaagcaacc
     7201 aggctggggt tagcgtgccg aggccatgtg gccccagcac ccggcacgat ctggcttggc
     7261 ggcgccgcgt tgccctgcct ccctaactag ggtgaggcca tcccgtccgg caccagttgc
     7321 gtgcgtggaa agatggccgc tcccgggccc tgttgcaagg agctcaaaat ggaggacgcg
     7381 gcagcccggt ggagcgggcg ggtgagtcac ccacacaaag gaagagggcc tggtccctca
     7441 ccggctgctg cttcctgtga ccccgt
//
