LOCUS       sequence-477262-        7940 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cytosolic expression of mTagBFP2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7940)
  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 7940)
  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 mTagBFP2
FEATURES             Location/Qualifiers
     source          1..7940
                     /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             3871..4581
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     misc_feature    4607..5195
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4660..4680)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5078..5089)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5198..5214)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5199..5215)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5720..5900
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5926..5943)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5932..6043
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(6189..6777)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6269..6288)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6948..7739,7740..7808))
                     /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     7571..7590
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7809..7913)
                     /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 atggtgtcta agggcgaaga gctgattaag
     3901 gagaacatgc acatgaagct gtacatggag ggcaccgtgg acaaccatca cttcaagtgc
     3961 acatccgagg gcgaaggcaa gccctacgag ggcacccaga ccatgagaat caaggtggtc
     4021 gagggcggcc ctctcccctt cgccttcgac atcctggcta ctagcttcct ctacggcagc
     4081 aagaccttca tcaaccacac ccagggcatc cccgacttct tcaagcagtc cttccctgag
     4141 ggcttcacat gggagagagt caccacatac gaagacgggg gcgtgctgac cgctacccag
     4201 gacaccagcc tccaggacgg ctgcctcatc tacaacgtca agatcagagg ggtgaacttc
     4261 acatccaacg gccctgtgat gcagaagaaa acactcggct gggaggcctt caccgagacg
     4321 ctgtaccccg ctgacggcgg cctggaaggc agaaacgaca tggccctgaa gctcgtgggc
     4381 gggagccatc tgatcgcaaa cgccaagacc acatatagat ccaagaaacc cgctaagaac
     4441 ctcaagatgc ctggcgtcta ctatgtggac tacagactgg aaagaatcaa ggaggccaac
     4501 aacgagacct acgtcgagca gcacgaggtg gcagtggcca gatactgcga cctccctagc
     4561 aaactggggc acaagcttaa tcgaattcga tatcaagctt atcgataatc aacctctgga
     4621 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     4681 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     4741 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     4801 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     4861 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     4921 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     4981 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     5041 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     5101 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     5161 gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgtcgac ctcgagacct
     5221 agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg
     5281 gctagaagca caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag
     5341 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact
     5401 ggaagggcta attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac
     5461 acaaggctac ttccctgatt ggcagaacta cacaccaggg ccagggatca gatatccact
     5521 gacctttgga tggtgctaca agctagtacc agttgagcaa gagaaggtag aagaagccaa
     5581 tgaaggagag aacacccgct tgttacaccc tgtgagcctg catgggatgg atgacccgga
     5641 gagagaagta ttagagtgga ggtttgacag ccgcctagca tttcatcaca tggcccgaga
     5701 gctgcatccg gactgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     5761 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     5821 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     5881 agtgtggaaa atctctagca gggcccgttt aaacccgctg atcagcctcg actgtgcctt
     5941 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     6001 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     6061 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     6121 atagcaggca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     6181 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     6241 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     6301 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     6361 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     6421 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6481 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6541 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6601 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     6661 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6721 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6781 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6841 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6901 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6961 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     7021 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     7081 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     7141 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     7201 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     7261 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     7321 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     7381 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     7441 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     7501 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7561 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7621 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7681 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7741 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7801 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7861 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7921 ccccgaaaag tgccacctga
//
