LOCUS       sequence-463818-        8762 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral construct of the MitoTRACER genetic reporter to be 
            expressed in the donor cells under the Synapsin1 promotor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8762)
  AUTHORS   Hoover G, Gilbert S, Curley O, Obellianne C, Lin MT, Hixson W, 
            Pierce TW, Andrews JF, Alexeyev MF, Ding Y, Bu P, Behbod F, Medina 
            D, Chang JT, Ayala G, Grelet S
  TITLE     Nerve-to-cancer transfer of mitochondria during cancer metastasis.
  JOURNAL   Nature. 2025 Jun 25. doi: 10.1038/s41586-025-09176-8.
  PUBMED    40562940
REFERENCE   2  (bases 1 to 8762)
  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: "Nature. 
            2025 Jun 25. doi: 10.1038/s41586-025-09176-8."
COMMENT     Sequence Label: [#GS16-LV] Syn1-MitoTRACER - Donor
FEATURES             Location/Qualifiers
     source          1..8762
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             736..1369
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    1416..1541
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2038..2271
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2456..2500
                     /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    2767..2884
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     regulatory      4077..4086
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4089..4109
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5139..5159
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             5178..5204
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5217..5264
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     misc_feature    5397..5985
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5450..5470)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5868..5879)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(6129..6162)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             6182..6362
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6381..6836)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6518..6539)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6730..6749
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6863..6967
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6969..7326
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6969..7160
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6992..7012)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      7177..7312
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             7361..8155
                     /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             7361..8155
                     /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(7415..7434)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8025..8044
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(8343..8362)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatatgtac
      721 tacaaactta gtagttggaa gggctaattc actcccaaag aagacaagat atccttgatc
      781 tgtggatcta ccacacacaa ggctacttcc ctgattagca gaactacaca ccagggccag
      841 gggtcagata tccactgacc tttggatggt gctacaagct agtaccagtt gagccagata
      901 aggtagaaga ggccaataaa ggagagaaca ccagcttgtt acaccctgtg agcctgcatg
      961 ggatggatga cccggagaga gaagtgttag agtggaggtt tgacagccgc ctagcatttc
     1021 atcacgtggc ccgagagctg catccggagt acttcaagaa ctgctgatat cgagcttgct
     1081 acaagggact ttccgctggg gactttccag ggaggcgtgg cctgggcggg actggggagt
     1141 ggcgagccct cagatcctgc atataagcag ctgctttttg cctgtactgg gtctctctgg
     1201 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     1261 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     1321 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga
     1381 acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg
     1441 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga
     1501 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa
     1561 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa
     1621 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag
     1681 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat
     1741 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga
     1801 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta
     1861 agaccaccgc acagcaagcg gccggccgct gatcttcaga cctggaggag gagatatgag
     1921 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
     1981 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
     2041 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac
     2101 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
     2161 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
     2221 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
     2281 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
     2341 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
     2401 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
     2461 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
     2521 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
     2581 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
     2641 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
     2701 aggtggagag agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg
     2761 gttaactttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
     2821 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa
     2881 ttttatcgat cacgagacta gcattaatgc atacccagag tttccttgct catctgggag
     2941 tctgtgtttt tcctagatgt gtgcacttgt gtgagattct ctgggtgtga gtcaaagtgt
     3001 tatctgaatg tgtaatgtgt gctcaatatg ctcatgtgtg ttaccctgag cttctgtgtc
     3061 tacatatata cctggatgcc tgtgtgttct gtgatgtaca tatatattct gtctttcctt
     3121 ccttttctat ttgtgttatt ccatgtgttc tttcagattc tcaccaccaa gggcaaggat
     3181 atgttaacta cccaagtgtc cacctccgcc tgtctggtga tgtttacgcc acccccgtgc
     3241 tcttttcttt gcccgacaga gttgttatag gagatgtctc cccgggaaca ctgcaggaag
     3301 gagaatttct acatttatgt tcccctctga gtgtgcttct atccccaaaa tgccttcaaa
     3361 ggtgaaaatc aacactggaa acccaagtat ctgggaaggg caagagtgtg taagtgcaag
     3421 ttagcctaag gaataggaag aggttggtaa acagggtagg atcgtgggag ggagtttcgt
     3481 tactacaggt ccggaccctc aggacaagaa ccccaccccc actccccaaa ttgcgcatcc
     3541 cccgccccca tcagaggggg aggggaagag gttgcggcgc ggcgcatgcg cactgtcgga
     3601 ttcagcaccg cggtcagagc cttcgcctcc gctgccggcg cgcaccacca cctccccagc
     3661 accaaaggct gactgacgcc actcactagc cctccccaaa ctccccttcc tcgccgcctt
     3721 ggtcgcgtcc atgctgccgt gagtccagtc ggaccgcacc acgagaggtg caagataggg
     3781 aggtgcgggc gcgaccatac gctctgcggc ggcagagcct cggcgctgcc tcagtctgca
     3841 gcgggcagca gaggagtcgc gtcgtgccag agagcaccgc cgtgctcctg agcccctcgc
     3901 gctccgcccc cgcggcccac agacccactg ccccttgggc tagctgttta aacctctaga
     3961 gctaccggtg cggccgcatt aattaactag gcgcgccaac tcgaccggac tcagatatcg
     4021 agctcaagct tcgaattctg cagtcgacgg taccgcgggc ccgggtacgg gaattcgcca
     4081 ccatggtgcc caagaagaag aggaaagtct ccaacctgct gactgtgcac caaaacctgc
     4141 ctgccctccc tgtggatgcc acctctgatg aagtcaggaa gaacctgatg gacatgttca
     4201 gggacaggca ggccttctct gaacacacct ggaagatgct cctgtctgtg tgcagatcct
     4261 gggctgcctg gtgcaagctg aacaacagga aatggttccc tgctgaacct gaggatgtga
     4321 gggactacct cctgtacctg caagccagag gcctggctgt gaagaccatc caacagcacc
     4381 tgggccagct caacatgctg cacaggagat ctggcctgcc tcgcccttct gactccaatg
     4441 ctgtgtccct ggtgatgagg agaatcagaa aggagaatgt ggatgctggg gagagagcca
     4501 agcaggccct ggcctttgaa cgcactgact ttgaccaagt cagatccctg atggagaact
     4561 ctgacagatg ccaggacatc aggaacctgg ccttcctggg cattgcctac aacaccctgc
     4621 tgcgcattgc cgaaattgcc agaatcagag tgaaggacat ctcccgcacc gatggtggga
     4681 gaatgctgat ccacattggc aggaccaaga ccctggtgtc cacagctggt gtggagaagg
     4741 ccctgtccct gggggttacc aagctggtgg agagatggat ctctgtgtct ggtgtggctg
     4801 atgaccccaa caactacctg ttctgccggg tcagaaagaa tggtgtggct gccccttctg
     4861 ccacctccca actgtccacc cgggccctgg aagggatctt tgaggccacc caccgcctga
     4921 tctatggtgc caaggatgac tctgggcaga gatacctggc ctggtctggc cactctgcca
     4981 gagtgggtgc tgccagggac atggccaggg ctggtgtgtc catccctgaa atcatgcagg
     5041 ctggtggctg gaccaatgtg aacattgtga tgaactacat cagaaacctg gactctgaga
     5101 ctggggccat ggtgaggctg ctagaggatg gggacagaga gaacctgtac ttccagtctc
     5161 tcgagggcag cgccggatac ccttatgatg tgcctgatta tgctggaggg agcggccgtg
     5221 atcatatggt tcttcatgaa tatgttaatg ctgctggtat tactccgcgg catcgaggcg
     5281 acggagagcc tagtggagtt cctgtagctg tggtgctgct gccagtgttt gcccttaccc
     5341 tggtagcagt ttgggccttc gtgagatacc gaaagcagct ctgagggaat tccgataatc
     5401 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     5461 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     5521 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     5581 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     5641 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     5701 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     5761 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct
     5821 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     5881 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     5941 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcgg gaattcgagc
     6001 tcggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     6061 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatgg gatcaattca
     6121 ccatgggaat aacttcgtat agcatacatt atacgaagtt atgctgcttt ttgcttgtac
     6181 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     6241 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     6301 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     6361 cagcatctag ctagcggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     6421 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     6481 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     6541 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     6601 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     6661 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     6721 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     6781 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     6841 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     6901 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     6961 gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     7021 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     7081 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     7141 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     7201 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     7261 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     7321 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     7381 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     7441 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     7501 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
     7561 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     7621 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     7681 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     7741 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     7801 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     7861 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
     7921 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     7981 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     8041 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     8101 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     8161 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     8221 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     8281 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
     8341 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     8401 cagaataaaa cgcgcggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     8461 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     8521 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     8581 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
     8641 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     8701 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     8761 tc
//
