LOCUS       sequence-236127-        5078 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for PRM-5R in a vector expressing 
            eGFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5078)
  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 5078)
  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-5R
FEATURES             Location/Qualifiers
     source          1..5078
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      21..30
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(27..29,30..32,33..746)
                     /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(27..29,30..32,33..743)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(72..93)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(333..352)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     680..701
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    869..990
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(906..925)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     960..979
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(997..1452)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1134..1155)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1346..1365
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1479..1583
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1585..1942
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1585..1776
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1608..1628)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1793..1928
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1977..2771
                     /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             1977..2771
                     /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(2031..2050)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2641..2660
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2959..2978)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3003..3050
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3379..3967
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3868..3887
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4142..4445
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4446..4649
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4600..4620
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4703..4732
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
ORIGIN
        1 gcggtggcgg ctcaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
       61 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
      121 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
      181 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
      241 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
      301 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
      361 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
      421 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
      481 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
      541 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
      601 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
      661 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
      721 tcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac
      781 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
      841 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
      901 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
      961 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt
     1021 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     1081 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     1141 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     1201 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     1261 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     1321 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     1381 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     1441 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     1501 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     1561 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     1621 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     1681 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     1741 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     1801 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     1861 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     1921 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     1981 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     2041 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     2101 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     2161 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     2221 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     2281 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     2341 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     2401 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     2461 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     2521 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     2581 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     2641 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     2701 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     2761 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     2821 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     2881 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     2941 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     3001 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     3061 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     3121 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     3181 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     3241 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     3301 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     3361 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3421 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3481 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3541 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3601 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3661 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3721 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3781 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3841 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3901 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3961 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4021 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     4081 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     4141 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     4201 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     4261 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     4321 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     4381 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     4441 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     4501 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     4561 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     4621 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct
     4681 accggactca gatctcgaga tggaacaaaa actcatctca gaagaggatc tgcgaattct
     4741 gcatatgaaa ggaggcagct gggggggtag cgcgccacct ccacctccac ctagccgtgg
     4801 gtggggctca ggtggttctg ggggtagcgg aggaagtgcg ccacctccac ctccacctag
     4861 ccgtggcggt ggcggctcag gtggttctgg gggtagcgga ggaagtgcgc cacctccacc
     4921 tccacctagc cgtggcggtg gcggctcacg ggatccaggt ggttctgggg gtagcggagg
     4981 aagtgcgcca cctccacctc cacctagccg tgggtggggc tcaggtggtt ctgggggtag
     5041 cggaggaagt gcgccacctc cacctccacc tagccgtg
//
