LOCUS       sequence-474175-        8298 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral construct of mitochondrial localized GFP 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 8298)
  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 8298)
  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: [#GS24 -LV] Syn1-GFP-OMP25
FEATURES             Location/Qualifiers
     source          1..8298
                     /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"
     misc_feature    4013..4069
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      4070..4079
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4076..4078,4079..4081,4082..4792)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4076..4078,4079..4081,4082..4792)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4121..4142)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4382..4401)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4729..4750
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4933..5521
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4986..5006)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5404..5415)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(5665..5698)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             5718..5898
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(5917..6372)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6054..6075)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6266..6285
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6399..6503
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6505..6862
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6505..6696
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6528..6548)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6713..6848
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6897..7691
                     /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             6897..7691
                     /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(6951..6970)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7561..7580
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7879..7898)
                     /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 ccgggatccg ccaccatggt
     4081 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     4141 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     4201 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     4261 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     4321 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     4381 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     4441 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     4501 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     4561 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     4621 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     4681 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     4741 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agccgcggca
     4801 tcgaggcgac ggagagccta gtggagttcc tgtagctgtg gtgctgctgc cagtgtttgc
     4861 ccttaccctg gtagcagttt gggccttcgt gagataccga aagcagctct gatagctcga
     4921 gggaattccg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     4981 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     5041 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     5101 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     5161 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     5221 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     5281 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     5341 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     5401 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     5461 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     5521 catcgggaat tcgagctcgg tacctttaag accaatgact tacaaggcag ctgtagatct
     5581 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
     5641 agatgggatc aattcaccat gggaataact tcgtatagca tacattatac gaagttatgc
     5701 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     5761 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     5821 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     5881 tgtggaaaat ctctagcagc atctagctag cggcgtaaat tgtaagcgtt aatattttgt
     5941 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     6001 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     6061 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     6121 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     6181 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     6241 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     6301 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     6361 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     6421 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     6481 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     6541 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     6601 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     6661 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     6721 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     6781 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     6841 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     6901 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     6961 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     7021 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     7081 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     7141 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     7201 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     7261 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     7321 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     7381 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     7441 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     7501 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     7561 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     7621 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     7681 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     7741 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     7801 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     7861 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     7921 gacggcaata aaaagacaga ataaaacgcg cggtgttggg tcgtttgttc ataaacgcgg
     7981 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     8041 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     8101 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     8161 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     8221 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     8281 aaagatcaaa ggatcttc
//
