LOCUS       sequence-362883-        7316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasma membrane-targeted mCardinal expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7316)
  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 7316)
  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-mCardinal-CAXX
FEATURES             Location/Qualifiers
     source          1..7316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        51..242
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(74..94)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        256..1562
                     /label=mEF-1-alpha promoter
                     /note="strong constitutive promoter for mouse elongation 
                     factor EF-1-alpha"
     intron          583..1530
                     /label=mEF-1-alpha intron
                     /note="intron upstream of the start codon of mouse 
                     EF-1-alpha"
     polyA_signal    1607..1728
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1618..1637)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1672..1691
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      1916..2504
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2405..2424
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2584..2887
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3002..4307
                     /label=rEF-1-alpha promoter
                     /note="strong constitutive promoter for rat elongation 
                     factor EF-1-alpha"
     misc_feature    3344..4275
                     /label=rEF-1-alpha intron
                     /note="intron upstream of the start codon of rat 
                     EF-1-alpha"
     regulatory      4314..4323
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4320..5051
                     /codon_start=1
                     /product="far-red fluorescent protein derived from 
                     mNeptune, with further red-shifted spectra"
                     /label=mCardinal
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTTEGEGKPYEGTQTQ
                     RIKVVEGGPLPFAFDILATCFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTVTQDTSLQDGCLIYNVKLRGVNFPSNGPVMQKKTLGWEATTETLYPADGGLEGRCD
                     MALKLVGGGHLHCNLKTTYRSKKPAKNLKMPGVYFVDRRLERIKEADNETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     misc_feature    5163..5607
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        5640..5686
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        5678..5696
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5706..6731
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    6743..7315
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
ORIGIN
        1 cctgcagggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga
       61 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
      121 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
      181 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
      241 ccactagtgg agccgagagt aattcataca aaaggaggga tcgccttcgc aaggggagag
      301 cccagggacc gtccctaaat tctcacagac ccaaatccct gtagccgccc cacgacagcg
      361 cgaggagcat gcgctcaggg ctgagcgcgg ggagagcaga gcacacaagc tcatagaccc
      421 tggtcgtggg ggggaggacc ggggagctgg cgcggggcaa actgggaaag cggtgtcgtg
      481 tgctggctcc gccctcttcc cgagggtggg ggagaacggt atataagtgc ggcagtcgcc
      541 ttggacgttc tttttcgcaa cgggtttgcc gtcagaacgc aggtgagggg cgggtgtggc
      601 ttccgcgggc cgccgagctg gaggtcctgc tccgagcggg ccgggccccg ctgtcgtcgg
      661 cggggattag ctgcgagcat tcccgcttcg agttgcgggc ggcgcgggag gcagagtgcg
      721 aggcctagcg gcaaccccgt agcctcgcct cgtgtccggc ttgaggccta gcgtggtgtc
      781 cgcgccgccg ccgcgtgcta ctccggccgc actctggtct tttttttttt tgttgttgtt
      841 gccctgctgc cttcgattgc cgttcagcaa taggggctaa caaagggagg gtgcggggct
      901 tgctcgcccg gagcccggag aggtcatggt tggggaggaa tggagggaca ggagtggcgg
      961 ctggggcccg cccgccttcg gagcacatgt ccgacgccac ctggatgggg cgaggcctgg
     1021 ggtttttccc gaagcaacca ggctggggtt agcgtgccga ggccatgtgg ccccagcacc
     1081 cggcacgatc tggcttggcg gcgccgcgtt gccctgcctc cctaactagg gtgaggccat
     1141 cccgtccggc accagttgcg tgcgtggaaa gatggccgct cccgggccct gttgcaagga
     1201 gctcaaaatg gaggacgcgg cagcccggtg gagcgggcgg gtgagtcacc cacacaaagg
     1261 aagagggcct ggtccctcac cggctgctgc ttcctgtgac cccgtggtcc tatcggccgc
     1321 aatagtcacc tcgggctttt gagcacggct agtcgcggcg gggggagggg atgtaatggc
     1381 gttggagttt gttcacattt ggtgggtgga gactagtcag gccagcctgg cgctggaagt
     1441 catttttgga atttgtcccc ttgagttttg agcggagcta attctcgggc ttcttagcgg
     1501 ttcaaaggta tcttttaaac ccttttttag gtgttgtgaa aaccaccgct aattcaaagc
     1561 aaccggtgat atcaaagatc ttgtacagct agctggccag acatgataag atacattgat
     1621 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     1681 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     1741 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     1801 aacctctaca aatgtggtat ggaaatgtta attaactagc catgaccaaa atcccttaac
     1861 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     1921 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     1981 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     2041 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     2101 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     2161 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     2221 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     2281 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     2341 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     2401 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     2461 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     2521 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttctta attaacctgc
     2581 aggcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     2641 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     2701 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     2761 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     2821 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     2881 taccatgatg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
     2941 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttgac tagtggagcc
     3001 gagagtaatt catacaaaag gagggatcgc cttcgcaagg ggagagccca gggaccgtcc
     3061 ctaaattctc acagacccaa atccctgtag ccgccccacg acagcgcgag gagcatgcgc
     3121 ccagggctga gcgcgggtag atcagagcac acaagctcac agtccccggc ggtgggggga
     3181 ggggcgcgct gagcgggggc cagggagctg gcgcggggca aactgggaaa gtggtgtcgt
     3241 gtgctggctc cgccctcttc ccgagggtgg gggagaacgg tatataagtg cggtagtcgc
     3301 cttggacgtt ctttttcgca acgggtttgc cgtcagaacg caggtgagtg gcgggtgtgg
     3361 cttccgcggg ccccggagct ggagccctgc tctgagcggg ccgggctgat atgcgagtgt
     3421 cgtccgcagg gtttagctgt gagcattccc acttcgagtg gcgggcggtg cgggggtgag
     3481 agtgcgaggc ctagcggcaa ccccgtagcc tcgcctcgtg tccggcttga ggcctagcgt
     3541 ggtgtccgcc gccgcgtgcc actccggccg cactatgcgt tttttgtcct tgctgccctc
     3601 gattgccttc cagcagcatg ggctaacaaa gggagggtgt ggggctcact cttaaggagc
     3661 ccatgaagct tacgttggat aggaatggaa gggcaggagg ggcgactggg gcccgcccgc
     3721 cttcggagca catgtccgac gccacctgga tggggcgagg cctgtggctt tccgaagcaa
     3781 tcgggcgtga gtttagccta cctgggccat gtggccctag cactgggcac ggtctggcct
     3841 ggcggtgccg cgttcccttg cctcccaaca agggtgaggc cgtcccgccc ggcaccagtt
     3901 gcttgcgcgg aaagatggcc gctcccgggg ccctgttgca aggagctcaa aatggaggac
     3961 gcggcagccc ggtggagcgg gcgggtgagt cacccacaca aaggaagagg gccttgcccc
     4021 tcgccggccg ctgcttcctg tgaccccgtg gtctatcggc cgcatagtca cctcgggctt
     4081 ctcttgagca ccgctcgtcg cggcgggggg aggggatcta atggcgttgg agtttgttca
     4141 catttggtgg gtggagacta gtcaggccag cctggcgctg gaagtcattc ttggaatttg
     4201 cccctttgag tttggagcga ggctaattct caagcctctt agcggttcaa aggtattttc
     4261 taaacccgtt tccaggtgtt gtgaaagcca ccgctaattc aaagcaatcc ggagccacca
     4321 tggtgagcaa gggcgaggag ctgatcaagg agaacatgca catgaagctg tacatggaag
     4381 gcaccgtgaa caaccaccac ttcaagtgca ccaccgaagg ggagggcaag ccctacgagg
     4441 gcacccagac ccagaggatt aaggtggtgg agggaggccc cctgccgttc gcattcgaca
     4501 tcctggccac ctgctttatg tacgggagca agaccttcat caaccacacc cagggcatcc
     4561 ccgatttctt taagcagtcc ttccctgagg gcttcacatg ggagagagtc accacatacg
     4621 aagacggggg cgtgcttacc gttacccagg acaccagcct ccaggacggc tgcttgatct
     4681 acaacgtcaa gctcagaggg gtgaacttcc catccaacgg ccctgtgatg cagaagaaaa
     4741 cactcggctg ggaggccacc accgagaccc tgtaccccgc tgacggcggc ctggaaggca
     4801 gatgcgacat ggccctgaag ctcgtgggcg ggggccacct gcactgcaac ctgaagacca
     4861 catacagatc caagaaaccc gctaagaacc tcaagatgcc cggcgtctac tttgtggacc
     4921 gcagactgga aagaatcaag gaggccgaca atgagaccta cgtcgagcag cacgaggtgg
     4981 ctgtggccag atactgcgac ctccctagca aactggggca caaacttaat ggcatggacg
     5041 agctgtacaa gtctggcagc ggaggctctg gaggcaaaga aaagatgagc aaagatggta
     5101 aaaagaagaa aaagaagtca aagacaaagt gtgtaattat gtgaggatcc cgtacgccta
     5161 ggagcaggtt tccccaatga cacaaaacgt gcaacttgaa actccgcctg gtctttccag
     5221 gtctagaggg gtaacacttt gtactgcgtt tggctccacg ctcgatccac tggcgagtgt
     5281 tagtaacagc actgttgctt cgtagcggag catgacggcc gtgggaactc ctccttggta
     5341 acaaggaccc acggggccaa aagccacgcc cacacgggcc cgtcatgtgt gcaaccccag
     5401 cacggcgact ttactgcgaa acccacttta aagtgacatt gaaactggta cccacacact
     5461 ggtgacaggc taaggatgcc cttcaggtac cccgaggtaa cacgcgacac tcgggatctg
     5521 agaaggggac tggggcttct ataaaagcgc tcggtttaaa aagcttctat gcctgaatag
     5581 gtgaccggag gtcggcacct ttcctttgca attactgacc ctatgaatac actgactgtt
     5641 tgacaattaa tcatcggcat agtatatcgg catagtataa tacgactcac tataggaggg
     5701 ccaccatgaa gaaacctgaa ctgacagcaa cttctgttga gaagtttctc attgaaaaat
     5761 ttgattctgt ttctgatctc atgcagctgt ctgaaggtga agaaagcaga gccttttctt
     5821 ttgatgttgg aggaagaggt tatgttctga gggtcaattc ttgtgctgat ggtttttaca
     5881 aagacagata tgtttacaga cactttgcct ctgctgctct gccaattcca gaagttctgg
     5941 acattggaga attttctgaa tctctcacct actgcatcag cagaagagca caaggagtca
     6001 ctctccagga tctccctgaa actgagctgc cagctgttct gcaacctgtt gctgaagcaa
     6061 tggatgccat tgcagcagct gatctgagcc aaacctctgg atttggtcct tttggtcccc
     6121 aaggcattgg tcagtacacc acttggaggg atttcatttg tgccattgct gatcctcatg
     6181 tctatcactg gcagactgtg atggatgaca cagtttctgc ttctgttgct caggcactgg
     6241 atgaactcat gctgtgggca gaagattgtc ctgaagtcag acacctggtc catgctgatt
     6301 ttggaagcaa caatgttctg acagacaatg gcagaatcac tgcagtcatt gactggtctg
     6361 aagccatgtt tggagattct caatatgagg ttgccaacat ttttttttgg agaccttggc
     6421 tggcttgcat ggaacaacaa acaagatatt ttgaaagaag acacccagaa ctggctggtt
     6481 cccccagact gagagcctac atgctcagaa ttggcctgga ccaactgtat caatctctgg
     6541 ttgatggaaa ctttgatgat gctgcttggg cacaaggaag atgtgatgcc attgtgaggt
     6601 ctggtgctgg aactgttgga agaactcaaa ttgcaagaag gtctgctgct gtttggactg
     6661 atggatgtgt tgaagttctg gctgactctg gaaacaggag accctccaca agacccagag
     6721 ccaaggaatg aatattagct agattatccc taatacctgc caccccactc ttaatcagtg
     6781 gtggaagaac ggtctcagaa ctgtttgttt caattggcca tttaagttta gtagtaaaag
     6841 actggttaat gataacaatg catcgtaaaa ccttcagaag gaaaggagaa tgttttgtgg
     6901 accactttgg ttttcttttt tgcgtgtggc agttttaagt tattagtttt taaaatcagt
     6961 actttttaat ggaaacaact tgaccaaaaa tttgtcacag aattttgaga cccattaaaa
     7021 aagttaaatg agaaacctgt gtgttccttt ggtcaacacc gagacattta ggtgaaagac
     7081 atctaattct ggttttacga atctggaaac ttcttgaaaa tgtaattctt gagttaacac
     7141 ttctgggtgg agaatagggt tgttttcccc ccacataatt ggaaggggaa ggaatatcat
     7201 ttaaagctat gggagggttg ctttgattac aacactggag agaaatgcag catgttgctg
     7261 attgcctgtc actaaaacag gccaaaaact gagtccttgg gttgcataga aagctg
//
