LOCUS       sequence-236132-        4919 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian eGFP expression plasmid for PRM-3R..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4919)
  AUTHORS   Li P, Banjade S, Cheng HC, Kim S, Chen B, Guo L, Llaguno M, 
            Hollingsworth JV, King DS, Banani SF, Russo PS, Jiang QX, Nixon BT, 
            Rosen MK
  TITLE     Phase transitions in the assembly of multivalent signalling 
            proteins.
  JOURNAL   Nature. 2012 Mar 7;483(7389):336-40. doi: 10.1038/nature10879.
  PUBMED    22398450
REFERENCE   2  (bases 1 to 4919)
  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"; journalName: 
            "Nature"; date: "2012-03-7- 7"; volume: "483"; issue: "7389"; pages:
            "336-40"
COMMENT     Sequence Label: eGFP-PRM-3R
FEATURES             Location/Qualifiers
     source          1..4919
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(184..203)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     531..552
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    720..841
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(757..776)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     811..830
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(848..1303)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(985..1006)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1197..1216
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1330..1434
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1436..1793
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1436..1627
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1459..1479)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1644..1779
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1828..2622
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1828..2622
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1882..1901)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2492..2511
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2810..2829)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2854..2901
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3230..3818
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3719..3738
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3993..4296
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4297..4500
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4451..4471
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4791..4800
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4797..4799,4800..4802,4803..4919,1..597)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4797..4799,4800..4802,4803..4919,1..594)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4842..4863)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
       61 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
      121 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
      181 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
      241 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
      301 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
      361 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
      421 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
      481 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
      541 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc
      601 ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa
      661 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
      721 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
      781 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
      841 aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
      901 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
      961 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     1021 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     1081 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     1141 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     1201 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     1261 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg
     1321 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     1381 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag
     1441 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     1501 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     1561 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     1621 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     1681 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     1741 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat
     1801 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     1861 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     1921 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     1981 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc
     2041 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     2101 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     2161 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     2221 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     2281 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     2341 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     2401 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     2461 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     2521 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     2581 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     2641 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     2701 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     2761 gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga
     2821 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     2881 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     2941 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     3001 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     3061 ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat ttttaattta
     3121 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     3181 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3241 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3301 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3361 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3421 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3481 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3541 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3601 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3661 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3721 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3781 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3841 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     3901 attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg
     3961 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     4021 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     4081 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     4141 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     4201 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     4261 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     4321 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     4381 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     4441 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     4501 ggtttagtga accgtcagat ccgctagcgc taccggactc agatctcgag ctcaagcttc
     4561 gaattctgca tatgaaagga ggcagctggg ggggtagcgc gccacctcca cctccaccta
     4621 gccgtgggtg gggctcaggt ggttctgggg gtagcggagg aagtgcgcca cctccacctc
     4681 cacctagccg tggcggtggc ggctcaggtg gttctggggg tagcggagga agtgcgccac
     4741 ctccacctcc acctagccgt ggcggtggcg gctcacggga tccaccggtc gccaccatgg
     4801 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     4861 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggc
//
