LOCUS       sequence-489751-        3600 bp    DNA     circular SYN 20-OCT-2025
DEFINITION  Expresses a + positively charged GFP variant (+15GFPKv) in mammalian
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3600)
  AUTHORS   Avecilla ARC, Thomas J, Quiroz FG
  TITLE     Genetically-Encoded Phase Separation Sensors Enable High-Fidelity 
            Live-Cell Probing of Biomolecular Condensates.
  JOURNAL   ACS Sens. 2025 Mar 28;10(3):1857-1869. doi: 
            10.1021/acssensors.4c02851. Epub 2025 Feb 23.
  PUBMED    39987501
REFERENCE   2  (bases 1 to 3600)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 20, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssensors.4c02851"; journalName: "ACS Sens"; date: 
            "2025-03-28- 28"; volume: "10"; issue: "3"; pages: "1857-1869"
COMMENT     Sequence Label: p15GFPkv
FEATURES             Location/Qualifiers
     source          1..3600
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          813..945
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      972..981
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1634..1655
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1773..1792)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    1802..1883
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     1827..1846
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(2081..2669)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2161..2180)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2750..3544)
                     /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             complement(2750..3544)
                     /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(2861..2880)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3471..3490
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagg
      721 tcgtttagtg aaccgtcaga tcactagtag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctc gactgatcac aggtaagtat caaggttaca agacaggttt
      841 aaggaggcca atagaaactg ggcttgtcga gacagagaag attcttgcgt ttctgatagg
      901 cacctattgg tcttactgac atccactttg cctttctctc cacaggggta ccgaagccgc
      961 tagcatctgt cgccaccatg ggcgcctcca agggcgagaa gctgttcacc ggcgtggtgc
     1021 ccatcctggt ggagctggac ggagacgtga acggccacaa attctccgtg cgcggcgagg
     1081 gcgagggcga cgccaccaaa ggcaagctga ccctgaagtt catctgcacc accgggaaac
     1141 tgcccgtgcc ctggcccacc ctggtgacca ccctgacata cggcgtgcag tgcttctccc
     1201 ggtatcccaa gcacatgaag cgccacgact tcttcaaatc cgccatgccc gagggctacg
     1261 tgcaggagcg caccatctcc ttcaagaagg acggcaccta caagacccgc gccgaggtga
     1321 agttcgaggg gaagaccctg gtgaaccgca tcgagctgaa gggaaaagac ttcaaggaga
     1381 agggcaacat cctgggccac aagctggagt acaacttcaa ctcccacaac gtgtatatca
     1441 cagccgacaa gaagaagaac ggcatcaagg ccaacttcaa gatccggcac aacgtgaaag
     1501 acggatctgt gcagctggcc gaccactacc agcagaacac ccccatcgga aaggggcccg
     1561 tgctgctgcc caagaaccac tacctgtcca caaaatccgc cctgtccaaa gaccccaagg
     1621 agaagcggga ccacatggtc ctgctggagt tcgtgacagc cgccgggatc acccacggca
     1681 tggacgagct gtacaagtca ggactggaac tcttggagga tctgaccctc ggcagccccg
     1741 ggtctccagg atcctaatga gaattcgagc tcgatgagtt tggacaaacc acaactagaa
     1801 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     1861 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     1921 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtacttaag
     1981 agggggagac caaagggcga gacgttaagg cctcacgtga catgtgagca aaaggccagc
     2041 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     2101 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     2161 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2221 cgcttacggg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     2281 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2341 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2401 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     2461 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     2521 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     2581 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     2641 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     2701 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgccgtct cagaagaact
     2761 cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca
     2821 cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg
     2881 ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc
     2941 ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct
     3001 cgccgtcggg catgctcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat
     3061 gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctctct
     3121 cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc
     3181 gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga
     3241 gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt
     3301 cgagtacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt
     3361 cttgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct
     3421 gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat
     3481 agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa
     3541 tcataatatt attgaagcat ttatcagggt tcgtctcgtc ccggtctcct cccatgcatg
//
