LOCUS       sequence-379566-        5400 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Bax and mitochodrial targeting sequence. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5400)
  AUTHORS   Scorrano L
  TITLE     Luca Scorrano plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 5400)
  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: 
            "unpublished"
COMMENT     Sequence Label: pcDNA3.1-HA-Bax-mito
FEATURES             Location/Qualifiers
     source          1..5400
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             137..931
                     /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             137..931
                     /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(191..210)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     801..820
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1119..1138)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1163..1210
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1539..2127
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2028..2047
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2302..2605
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2606..2809
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2760..2780
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2838..2912
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     regulatory      2937..2946
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2949..2975
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      3581..3590
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3587..3589,3590..3592,3593..4306)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3632..3653)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3893..3912)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4240..4261
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4429..4550
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4466..4485)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4520..4539
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4557..5012)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4694..4715)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4906..4925
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5039..5143
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(5145..5400,1..102)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5145..5336
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5168..5188)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(5353..5400,1..88)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
       61 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
      121 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
      181 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
      241 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
      301 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
      361 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
      421 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
      481 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
      541 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
      601 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
      661 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
      721 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
      781 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
      841 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
      901 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
      961 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     1021 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     1081 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     1141 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     1201 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     1261 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     1321 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     1381 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     1441 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     1501 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1561 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     1621 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     1681 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     1741 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     1801 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     1861 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     1921 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     1981 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2041 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2101 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2161 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     2221 aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt cattagttca
     2281 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     2341 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     2401 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     2461 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     2521 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     2581 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     2641 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     2701 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     2761 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     2821 ccgtcagatc cgctagcatg tccgtcctga cgccgctgct gctgcggggc ttgacaggct
     2881 cggcccggcg gctcccagtg ccgcgcgcca agatccattc gttgggggat caattcgcca
     2941 ccatggcata cccatacgac gtcccagact acgctatgga cgggtccggg gagcagcttg
     3001 ggagcggcgg gcccaccagc tctgaacaga tcatgaagac aggggccttt ttgctacagg
     3061 gtttcatcca ggatcgagca gggaggatgg ctggggagac acctgagctg accttggagc
     3121 agccgcccca ggatgcgtcc accaagaagc tgagcgagtg tctccggcga attggagatg
     3181 aactggacag caatatggag ctgcagagga tgattgctga cgtggacacg gactcccccc
     3241 gagaggtctt cttccgggtg gcagctgaca tgtttgctga tggcaacttc aactggggcc
     3301 gcgtggttgc cctcttctac tttgctagca aactggtgct caaggccctg tgcactaaag
     3361 tgcccgagct gatcagaacc atcatgggct ggacactgga cttcctccgt gagcggctgc
     3421 ttgtctggat ccaagaccag ggtggctggg aaggcctcct ctcctacttc gggaccccca
     3481 catggcagac agtgaccatc tttgtggctg gagtcctcac cgcctcgctc accatctgga
     3541 agaagatggg ctgagaataa gccgaattga tccaccggtc gccaccatgg tgagcaaggg
     3601 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     3661 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     3721 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     3781 gacctggggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     3841 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     3901 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     3961 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     4021 ctacatcagc cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa
     4081 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     4141 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     4201 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     4261 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgcgactc
     4321 tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     4381 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     4441 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4501 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat
     4561 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     4621 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     4681 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     4741 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     4801 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     4861 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     4921 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     4981 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     5041 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     5101 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     5161 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     5221 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     5281 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     5341 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
//
