LOCUS       sequence-469990-        6526 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express YFP in mitochondria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6526)
  AUTHORS   Ernst P, Xu N, Qu J, Chen H, Goldberg MS, Darley-Usmar V, Zhang JJ, 
            O'Rourke B, Liu X, Zhou L
  TITLE     Precisely Control Mitochondria with Light to Manipulate Cell Fate 
            Decision.
  JOURNAL   Biophys J. 2019 Aug 20;117(4):631-645. doi: 
            10.1016/j.bpj.2019.06.038. Epub 2019 Jul 26.
  PUBMED    31400914
REFERENCE   2  (bases 1 to 6526)
  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: 
            "10.1016/j.bpj.2019.06.038"; journalName: "Biophys J"; date: 
            "2019-08-20- 20"; volume: "117"; issue: "4"; pages: "631-645"
COMMENT     Sequence Label: pCMV-ABCB10-eYFP
FEATURES             Location/Qualifiers
     source          1..6526
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(1321..1323,1324..1326,1327..2037)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1366..1387)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1627..1646)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1974..1995
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(2079..2097)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2080..2097)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2117..2134)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2123..2347
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2393..2821
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2480..2499)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2690..2711
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2830..2850)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2835..3164
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3015..3150
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3077..3096
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3231..4025
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3285..3304)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3895..3914
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4199..4320
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4236..4255)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4290..4309
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4369..4385)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4369..4385)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4382..4404)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4393..4409
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4417..4423,4424..4441,4442..4447))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4462..4483
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4600..4617)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4771..5359)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4851..4870)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5530..6321,6322..6390))
                     /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     6153..6172
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6391..6495)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgccacc
      901 atgcgcgccc cttctgctag ggcgctactg ctgattccgc gtcggggccc tgccgtgcga
      961 gcgtgggccc cggccgtctc ctctcggata tggctggctt ctgaatggac cccgctcgta
     1021 cgcgcgtgga cctctctgat ccacaagccg ggttcgggcc tccgctttcc cgcgccccta
     1081 tccgggctgc ctggcggcgt ggggcagtgg gccacctcct cgggggcccg caggtgctgg
     1141 gtgctggcag gaccccgcgc cgcacatccc ctgttcgcca ggctccaggg tgcagctgcc
     1201 accggtgtgc gagaccttgg gaacgactcg cagcggcgtc ccgcggcgac cgggcgctca
     1261 gaagtatgga agctcctagg gctggtgcgc cccgagcgcg ggagactgtc agctgcagtt
     1321 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     1381 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     1441 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     1501 ctcgtgacca ccttcggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag
     1561 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     1621 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     1681 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     1741 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     1801 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     1861 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     1921 tacctgagct accagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     1981 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagttc
     2041 tgctacgaga acgaggtgta aggcgcgccg ggccctattc tatagtgtca cctaaatgct
     2101 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     2161 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     2221 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     2281 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     2341 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     2401 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2461 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2521 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2581 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2641 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2701 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2761 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2821 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     2881 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     2941 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3001 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3061 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     3121 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     3181 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3241 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3301 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3361 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     3421 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3481 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3541 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3601 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3661 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3721 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     3781 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     3841 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     3901 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     3961 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4021 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4081 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4141 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4201 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4261 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4321 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     4381 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4441 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4501 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4561 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4621 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4681 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4741 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4801 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4861 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4921 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4981 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5041 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5101 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5161 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5221 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5281 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5341 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5401 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5461 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5521 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5581 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5641 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5701 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5761 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5821 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5881 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5941 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6001 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6061 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6121 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6181 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6241 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6301 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6361 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6421 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6481 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
