LOCUS       sequence-477261-        7955 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cytosolic expression of mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7955)
  AUTHORS   Goldman, C., Kareva, T., Sarrafha, L., Schuldt, B.R., Sahasrabudhe, 
            A., Ahfeldt, T.,, Blanchard, J.W
  TITLE     Genetically encoded and modular subcellular organelle probes reveal 
            dysfunction in lysosomes and mitochondria driven by PRKN knockout
  JOURNAL   iScience 28 (2025)
REFERENCE   2  (bases 1 to 7955)
  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"; journalName: "iScience 28
            (2025)"
COMMENT     Sequence Label: pFUW mEmerald
FEATURES             Location/Qualifiers
     source          1..7955
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(48..67)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        239..618
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        620..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     773..793
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             836..1016
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1063..1188
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1681..1914
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2099..2143
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2441..2558
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2623..3834
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3671..3690
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      3865..3874
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3871..3873,3874..3876,3877..4590)
                     /codon_start=1
                     /product="brightly fluorescent monomeric variant of GFP"
                     /label=mEmerald
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3916..3937)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4177..4196)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4524..4545
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4622..5210
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4675..4695)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5093..5104)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5213..5229)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5214..5230)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5735..5915
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5941..5958)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5947..6058
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(6204..6792)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6284..6303)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6963..7754,7755..7823))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7586..7605
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7824..7928)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
       61 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
      121 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
      181 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
      241 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      301 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      361 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      421 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      481 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      541 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      601 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      661 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      721 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      781 ggcggtaggc gtgtacggtg ggaggtctat ataagcagcg cgttttgcct gtactgggtc
      841 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
      901 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
      961 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
     1021 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
     1081 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
     1141 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
     1201 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
     1261 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
     1321 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
     1381 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
     1441 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
     1501 aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat
     1561 gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg
     1621 agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat
     1681 aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat
     1741 gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt
     1801 gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca
     1861 gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat
     1921 ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag
     1981 taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat
     2041 taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa
     2101 gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat
     2161 aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt
     2221 aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt
     2281 atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga
     2341 agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat cggcactgcg
     2401 tgcgccaatt ctgcagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
     2461 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
     2521 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
     2581 gagatccagt ttggttaatt aacccgtgtc ggctccagat ctggcctccg cgccgggttt
     2641 tggcgcctcc cgcgggcgcc cccctcctca cggcgagcgc tgccacgtca gacgaagggc
     2701 gcagcgagcg tcctgatcct tccgcccgga cgctcaggac agcggcccgc tgctcataag
     2761 actcggcctt agaaccccag tatcagcaga aggacatttt aggacgggac ttgggtgact
     2821 ctagggcact ggttttcttt ccagagagcg gaacaggcga ggaaaagtag tcccttctcg
     2881 gcgattctgc ggagggatct ccgtggggcg gtgaacgccg atgattatat aaggacgcgc
     2941 cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg
     3001 atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg ctttcgtggc
     3061 cgccgggccg ctcggtggga cggaagcgtg tggagagacc gccaagggct gtagtctggg
     3121 tccgcgagca aggttgccct gaactggggg ttggggggag cgcagcaaaa tggcggctgt
     3181 tcccgagtct tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg aaacaaggtg
     3241 gggggcatgg tgggcggcaa gaacccaagg tcttgaggcc ttcgctaatg cgggaaagct
     3301 cttattcggg tgagatgggc tggggcacca tctggggacc ctgacgtgaa gtttgtcact
     3361 gactggagaa ctcggtttgt cgtctgttgc gggggcggca gttatggcgg tgccgttggg
     3421 cagtgcaccc gtacctttgg gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg
     3481 ttggcttata atgcagggtg gggccacctg ccggtaggtg tgcggtaggc ttttctccgt
     3541 cgcaggacgc agggttcggg cctagggtag gctctcctga atcgacaggc gccggacctc
     3601 tggtgagggg agggataagt gaggcgtcag tttctttggt cggttttatg tacctatctt
     3661 cttaagtagc tgaagctccg gttttgaact atgcgctcgg ggttggcgag tgtgttttgt
     3721 gaagtttttt aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt
     3781 tagactagta aattgtccgc taaattctgg ccgtttttgg cttttttgtt agacgaagct
     3841 tgggcccggg aattaattat taaggccacc atggtgagca agggcgagga gctgttcacc
     3901 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     3961 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     4021 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttgaccta cggcgtgcag
     4081 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     4141 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     4201 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     4261 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaag
     4321 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg tgaacttcaa gacccgccac
     4381 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     4441 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccaa gctgagcaaa
     4501 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     4561 actctcggca tggacgagct gtacaagtaa cggatccgaa ttcgatatca agcttatcga
     4621 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     4681 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     4741 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     4801 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     4861 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     4921 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     4981 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     5041 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     5101 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     5161 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     5221 tcgacctcga gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa
     5281 tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc
     5341 tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     5401 agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagata tccttgatct
     5461 gtggatctac cacacacaag gctacttccc tgattggcag aactacacac cagggccagg
     5521 gatcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg agcaagagaa
     5581 ggtagaagaa gccaatgaag gagagaacac ccgcttgtta caccctgtga gcctgcatgg
     5641 gatggatgac ccggagagag aagtattaga gtggaggttt gacagccgcc tagcatttca
     5701 tcacatggcc cgagagctgc atccggactg tactgggtct ctctggttag accagatctg
     5761 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     5821 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     5881 cagacccttt tagtcagtgt ggaaaatctc tagcagggcc cgtttaaacc cgctgatcag
     5941 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     6001 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     6061 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     6121 aggattggga agacaatagc aggcatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6181 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6241 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6301 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6361 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6421 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6481 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6541 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6601 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     6661 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6721 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6781 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6841 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6901 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6961 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7021 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7081 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7141 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7201 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7261 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7321 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7381 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7441 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7501 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7561 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7621 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7681 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7741 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7801 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7861 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7921 gttccgcgca catttccccg aaaagtgcca cctga
//
