LOCUS       sequence-362892-        8993 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nuclear membrane-targeted EBFP2 expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8993)
  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 8993)
  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-BFP-LaminB1
FEATURES             Location/Qualifiers
     source          1..8993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        236..427
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(259..279)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        441..1747
                     /label=mEF-1-alpha promoter
                     /note="strong constitutive promoter for mouse elongation 
                     factor EF-1-alpha"
     intron          768..1715
                     /label=mEF-1-alpha intron
                     /note="intron upstream of the start codon of mouse 
                     EF-1-alpha"
     polyA_signal    1792..1913
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1803..1822)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1857..1876
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2101..2689
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2590..2609
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2769..3072
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3187..4492
                     /label=rEF-1-alpha promoter
                     /note="strong constitutive promoter for rat elongation 
                     factor EF-1-alpha"
     misc_feature    3529..4460
                     /label=rEF-1-alpha intron
                     /note="intron upstream of the start codon of rat 
                     EF-1-alpha"
     regulatory      4499..4508
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4505..4507,4508..4510,4511..5221)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(4550..4571)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    7025..7469
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        7502..7548
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        7540..7558
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7568..8593
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    join(8605..8993,1..184)
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
ORIGIN
        1 agttaacact tctgggtgga gaatagggtt gttttccccc cacataattg gaaggggaag
       61 gaatatcatt taaagctatg ggagggttgc tttgattaca acactggaga gaaatgcagc
      121 atgttgctga ttgcctgtca ctaaaacagg ccaaaaactg agtccttggg ttgcatagaa
      181 agctgcctgc agggcctgaa ataacctctg aaagaggaac ttggttaggt accttctgag
      241 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
      301 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
      361 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
      421 tagtcccact agtggagccg agagtaattc atacaaaagg agggatcgcc ttcgcaaggg
      481 gagagcccag ggaccgtccc taaattctca cagacccaaa tccctgtagc cgccccacga
      541 cagcgcgagg agcatgcgct cagggctgag cgcggggaga gcagagcaca caagctcata
      601 gaccctggtc gtggggggga ggaccgggga gctggcgcgg ggcaaactgg gaaagcggtg
      661 tcgtgtgctg gctccgccct cttcccgagg gtgggggaga acggtatata agtgcggcag
      721 tcgccttgga cgttcttttt cgcaacgggt ttgccgtcag aacgcaggtg aggggcgggt
      781 gtggcttccg cgggccgccg agctggaggt cctgctccga gcgggccggg ccccgctgtc
      841 gtcggcgggg attagctgcg agcattcccg cttcgagttg cgggcggcgc gggaggcaga
      901 gtgcgaggcc tagcggcaac cccgtagcct cgcctcgtgt ccggcttgag gcctagcgtg
      961 gtgtccgcgc cgccgccgcg tgctactccg gccgcactct ggtctttttt ttttttgttg
     1021 ttgttgccct gctgccttcg attgccgttc agcaataggg gctaacaaag ggagggtgcg
     1081 gggcttgctc gcccggagcc cggagaggtc atggttgggg aggaatggag ggacaggagt
     1141 ggcggctggg gcccgcccgc cttcggagca catgtccgac gccacctgga tggggcgagg
     1201 cctggggttt ttcccgaagc aaccaggctg gggttagcgt gccgaggcca tgtggcccca
     1261 gcacccggca cgatctggct tggcggcgcc gcgttgccct gcctccctaa ctagggtgag
     1321 gccatcccgt ccggcaccag ttgcgtgcgt ggaaagatgg ccgctcccgg gccctgttgc
     1381 aaggagctca aaatggagga cgcggcagcc cggtggagcg ggcgggtgag tcacccacac
     1441 aaaggaagag ggcctggtcc ctcaccggct gctgcttcct gtgaccccgt ggtcctatcg
     1501 gccgcaatag tcacctcggg cttttgagca cggctagtcg cggcgggggg aggggatgta
     1561 atggcgttgg agtttgttca catttggtgg gtggagacta gtcaggccag cctggcgctg
     1621 gaagtcattt ttggaatttg tccccttgag ttttgagcgg agctaattct cgggcttctt
     1681 agcggttcaa aggtatcttt taaacccttt tttaggtgtt gtgaaaacca ccgctaattc
     1741 aaagcaaccg gtgatatcaa agatcttgta cagctagctg gccagacatg ataagataca
     1801 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     1861 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     1921 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     1981 agtaaaacct ctacaaatgt ggtatggaaa tgttaattaa ctagccatga ccaaaatccc
     2041 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2101 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2161 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2221 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     2281 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2341 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2401 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2461 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2521 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2581 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2641 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2701 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tcttaattaa
     2761 cctgcaggcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     2821 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     2881 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     2941 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     3001 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     3061 gctattacca tgatgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     3121 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttgactagtg
     3181 gagccgagag taattcatac aaaaggaggg atcgccttcg caaggggaga gcccagggac
     3241 cgtccctaaa ttctcacaga cccaaatccc tgtagccgcc ccacgacagc gcgaggagca
     3301 tgcgcccagg gctgagcgcg ggtagatcag agcacacaag ctcacagtcc ccggcggtgg
     3361 ggggaggggc gcgctgagcg ggggccaggg agctggcgcg gggcaaactg ggaaagtggt
     3421 gtcgtgtgct ggctccgccc tcttcccgag ggtgggggag aacggtatat aagtgcggta
     3481 gtcgccttgg acgttctttt tcgcaacggg tttgccgtca gaacgcaggt gagtggcggg
     3541 tgtggcttcc gcgggccccg gagctggagc cctgctctga gcgggccggg ctgatatgcg
     3601 agtgtcgtcc gcagggttta gctgtgagca ttcccacttc gagtggcggg cggtgcgggg
     3661 gtgagagtgc gaggcctagc ggcaaccccg tagcctcgcc tcgtgtccgg cttgaggcct
     3721 agcgtggtgt ccgccgccgc gtgccactcc ggccgcacta tgcgtttttt gtccttgctg
     3781 ccctcgattg ccttccagca gcatgggcta acaaagggag ggtgtggggc tcactcttaa
     3841 ggagcccatg aagcttacgt tggataggaa tggaagggca ggaggggcga ctggggcccg
     3901 cccgccttcg gagcacatgt ccgacgccac ctggatgggg cgaggcctgt ggctttccga
     3961 agcaatcggg cgtgagttta gcctacctgg gccatgtggc cctagcactg ggcacggtct
     4021 ggcctggcgg tgccgcgttc ccttgcctcc caacaagggt gaggccgtcc cgcccggcac
     4081 cagttgcttg cgcggaaaga tggccgctcc cggggccctg ttgcaaggag ctcaaaatgg
     4141 aggacgcggc agcccggtgg agcgggcggg tgagtcaccc acacaaagga agagggcctt
     4201 gcccctcgcc ggccgctgct tcctgtgacc ccgtggtcta tcggccgcat agtcacctcg
     4261 ggcttctctt gagcaccgct cgtcgcggcg gggggagggg atctaatggc gttggagttt
     4321 gttcacattt ggtgggtgga gactagtcag gccagcctgg cgctggaagt cattcttgga
     4381 atttgcccct ttgagtttgg agcgaggcta attctcaagc ctcttagcgg ttcaaaggta
     4441 ttttctaaac ccgtttccag gtgttgtgaa agccaccgct aattcaaagc aatccggagc
     4501 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     4561 ggacggcgac gtaaacggcc acaagttcag cgtgaggggc gagggcgagg gcgatgccac
     4621 caacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     4681 caccctcgtg accaccctga gccacggcgt gcagtgcttc gcccgctacc ccgaccacat
     4741 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     4801 cttcttcaag gacgacggca cctacaagac ccgcgccgag gtgaagttcg agggcgacac
     4861 cctggtgaac cgcatcgagc tgaagggcgt cgacttcaag gaggacggca acatcctggg
     4921 gcacaagctg gagtacaact tcaacagcca caacatctat atcatggccg tcaagcagaa
     4981 gaacggcatc aaggtgaact tcaagatccg ccacaacgtg gaggacggca gcgtgcagct
     5041 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacag
     5101 ccactacctg agcacccagt ccgtgctgag caaagacccc aacgagaagc gcgatcacat
     5161 ggtcctgctg gagttccgca ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     5221 gtctggcagc ggaggctctg gaggcatggc gactgcgacc cccgtgccgc cgcggatggg
     5281 cagccgcgct ggcggcccca ccacgccgct gagccccacg cgcctgtcgc ggctccagga
     5341 gaaggaggag ctgcgcgagc tcaatgaccg gctggcggtg tacatcgaca aggtgcgcag
     5401 cctggagacg gagaacagcg cgctgcagct gcaggtgacg gagcgcgagg aggtgcgcgg
     5461 ccgtgagctc accggcctca aggcgctcta cgagaccgag ctggccgacg cgcgacgcgc
     5521 gctcgacgac acggcccgcg agcgcgccaa gctgcagatc gagctgggca agtgcaaggc
     5581 ggaacacgac cagctgctcc tcaactatgc taagaaggaa tctgatctta atggcgccca
     5641 gatcaagctt cgagaatatg aagcagcact caattcgaaa gatgcagctc ttgctactgc
     5701 acttggtgac aaaaaaagtt tagagggaga tttggaggat ctgaaggatc agattgccca
     5761 gttggaagcc tccttagctg cagccaaaaa acagttagca gatgaaactt tacttaaagt
     5821 agatttggag aatcgttgtc agagccttac tgaggacttg gagtttcgca aaagcatgta
     5881 tgaagaggag attaacgaga ccagaaggaa gcatgaaacg cgcttggtag aggtggattc
     5941 tgggcgtcaa attgagtatg agtacaagct ggcgcaagcc cttcatgaga tgagagagca
     6001 acatgatgcc caagtgaggc tgtataagga ggagctggag cagacttacc atgccaaact
     6061 tgagaatgcc agactgtcat cagagatgaa tacttctact gtcaacagtg ccagggaaga
     6121 actgatggaa agccgcatga gaattgagag cctttcatcc cagctttcta atctacagaa
     6181 agagtctaga gcatgtttgg aaaggattca agaattagag gacttgcttg ctaaagaaaa
     6241 agacaactct cgtcgcatgc tgacagacaa agagagagag atggcggaaa taagggatca
     6301 aatgcagcaa cagctgaatg actatgaaca gcttcttgat gtaaagttag ccctggacat
     6361 ggaaatcagt gcttacagga aactcttaga aggcgaagaa gagaggttga agctgtctcc
     6421 aagcccttct tcccgtgtga cagtatcccg agcatcctca agtcgtagtg tacgtacaac
     6481 tagaggaaag cggaagaggg ttgatgtgga agaatcagag gcgagtagta gtgttagcat
     6541 ctctcattcc gcctcagcca ctggaaatgt ttgcatcgaa gaaattgatg ttgatgggaa
     6601 atttatccgc ttgaagaaca cttctgaaca ggatcaacca atgggaggct gggagatgat
     6661 cagaaaaatt ggagacacat cagtcagtta taaatatacc tcaagatatg tgctgaaggc
     6721 aggccagact gttacaattt gggctgcaaa cgctggtgtc acagccagcc ccccaactga
     6781 cctcatctgg aagaaccaga actcgtgggg cactggcgaa gatgtgaagg ttatattgaa
     6841 aaattctcag ggagaggagg ttgctcaaag aagtacagtc tttaaaacaa ccatacctga
     6901 agaagaggag gaggaggaag aagcagctgg agtggttgtt gaggaagaac ttttccacca
     6961 gcagggaacc ccaagagcat ccaatagaag ctgtgcaatt atgtgaggat cccgtacgcc
     7021 taggagcagg tttccccaat gacacaaaac gtgcaacttg aaactccgcc tggtctttcc
     7081 aggtctagag gggtaacact ttgtactgcg tttggctcca cgctcgatcc actggcgagt
     7141 gttagtaaca gcactgttgc ttcgtagcgg agcatgacgg ccgtgggaac tcctccttgg
     7201 taacaaggac ccacggggcc aaaagccacg cccacacggg cccgtcatgt gtgcaacccc
     7261 agcacggcga ctttactgcg aaacccactt taaagtgaca ttgaaactgg tacccacaca
     7321 ctggtgacag gctaaggatg cccttcaggt accccgaggt aacacgcgac actcgggatc
     7381 tgagaagggg actggggctt ctataaaagc gctcggttta aaaagcttct atgcctgaat
     7441 aggtgaccgg aggtcggcac ctttcctttg caattactga ccctatgaat acactgactg
     7501 tttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgactc actataggag
     7561 ggccaccatg aagaaacctg aactgacagc aacttctgtt gagaagtttc tcattgaaaa
     7621 atttgattct gtttctgatc tcatgcagct gtctgaaggt gaagaaagca gagccttttc
     7681 ttttgatgtt ggaggaagag gttatgttct gagggtcaat tcttgtgctg atggttttta
     7741 caaagacaga tatgtttaca gacactttgc ctctgctgct ctgccaattc cagaagttct
     7801 ggacattgga gaattttctg aatctctcac ctactgcatc agcagaagag cacaaggagt
     7861 cactctccag gatctccctg aaactgagct gccagctgtt ctgcaacctg ttgctgaagc
     7921 aatggatgcc attgcagcag ctgatctgag ccaaacctct ggatttggtc cttttggtcc
     7981 ccaaggcatt ggtcagtaca ccacttggag ggatttcatt tgtgccattg ctgatcctca
     8041 tgtctatcac tggcagactg tgatggatga cacagtttct gcttctgttg ctcaggcact
     8101 ggatgaactc atgctgtggg cagaagattg tcctgaagtc agacacctgg tccatgctga
     8161 ttttggaagc aacaatgttc tgacagacaa tggcagaatc actgcagtca ttgactggtc
     8221 tgaagccatg tttggagatt ctcaatatga ggttgccaac attttttttt ggagaccttg
     8281 gctggcttgc atggaacaac aaacaagata ttttgaaaga agacacccag aactggctgg
     8341 ttcccccaga ctgagagcct acatgctcag aattggcctg gaccaactgt atcaatctct
     8401 ggttgatgga aactttgatg atgctgcttg ggcacaagga agatgtgatg ccattgtgag
     8461 gtctggtgct ggaactgttg gaagaactca aattgcaaga aggtctgctg ctgtttggac
     8521 tgatggatgt gttgaagttc tggctgactc tggaaacagg agaccctcca caagacccag
     8581 agccaaggaa tgaatattag ctagattatc cctaatacct gccaccccac tcttaatcag
     8641 tggtggaaga acggtctcag aactgtttgt ttcaattggc catttaagtt tagtagtaaa
     8701 agactggtta atgataacaa tgcatcgtaa aaccttcaga aggaaaggag aatgttttgt
     8761 ggaccacttt ggttttcttt tttgcgtgtg gcagttttaa gttattagtt tttaaaatca
     8821 gtacttttta atggaaacaa cttgaccaaa aatttgtcac agaattttga gacccattaa
     8881 aaaagttaaa tgagaaacct gtgtgttcct ttggtcaaca ccgagacatt taggtgaaag
     8941 acatctaatt ctggttttac gaatctggaa acttcttgaa aatgtaattc ttg
//
