LOCUS       sequence-465303-        5627 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes a GFP-tagged Prohibitin2/PHB2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5627)
  AUTHORS   Bertolin G, Alves-Guerra MC, Cheron A, Burel A, Prigent C, Le Borgne
            R, Tramier M
  TITLE     Mitochondrial Aurora kinase A induces mitophagy by interacting with 
            MAP1LC3 and Prohibitin 2.
  JOURNAL   Life Sci Alliance. 2021 Apr 5;4(6):e202000806. doi: 
            10.26508/lsa.202000806. Print 2021 Jun.
  PUBMED    33820826
REFERENCE   2  (bases 1 to 5627)
  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: "Life Sci 
            Alliance."; date: "2021-04-5"; volume: "4(6):e202000806. doi"; 
            pages: " 10.26508/lsa.202000806. Print 2021 Jun"
COMMENT     Sequence Label: PHB2-GFP
FEATURES             Location/Qualifiers
     source          1..5627
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     303..322
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(621..640)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    665..712
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1041..1629
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1530..1549
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1804..2107
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2108..2311
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2262..2282
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3310..3319
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3316..3318,3319..3321,3322..4035)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3316..3318,3319..3321,3322..4032)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3361..3382)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3622..3641)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3969..3990
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4158..4279
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4195..4214)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4249..4268
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4286..4741)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4423..4444)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4635..4654
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4768..4872
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4874..5231
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4874..5065
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4897..4917)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5082..5217
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(5266..5627,1..433)
                     /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             join(5266..5627,1..433)
                     /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(5320..5339)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
       61 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
      121 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
      181 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
      241 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
      301 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
      361 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
      421 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
      481 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
      541 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
      601 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
      661 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
      721 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
      781 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
      841 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
      901 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
      961 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     1021 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     1081 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     1141 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     1201 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     1261 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     1321 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     1381 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     1441 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     1501 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     1561 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     1621 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     1681 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     1741 cattagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     1801 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     1861 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     1921 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     1981 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     2041 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     2101 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
     2161 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     2221 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     2281 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg
     2341 ctaccggact cagatctcga gatggcccag aacttgaagg acttggcggg acggctgccc
     2401 gccgggcccc ggggcatggg cacggccctg aagctgttgc tgggggccgg cgccgtggcc
     2461 tacggtgtgc gcgaatctgt gttcaccgtg gaaggcgggc acagagccat cttcttcaat
     2521 cggatcggtg gagtgcagca ggacactatc ctggccgagg gccttcactt caggatccct
     2581 tggttccagt accccattat ctatgacatt cgggccagac ctcgaaaaat ctcctcccct
     2641 acaggctcca aagacctaca gatggtgaat atctccctgc gagtgttgtc tcgacccaat
     2701 gctcaggagc ttcctagcat gtaccagcgc ctagggctgg actacgagga acgagtgttg
     2761 ccgtccattg tcaacgaggt gctcaagagt gtggtggcca agttcaatgc ctcacagctg
     2821 atcacccagc gggcccaggt atccctgttg atccgccggg agctgacaga gagggccaag
     2881 gacttcagcc tcatcctgga tgatgtggcc atcacagagc tgagctttag ccgagagtac
     2941 acagctgctg tagaagccaa acaagtggcc cagcaggagg cccagcgggc ccaattcttg
     3001 gtagaaaaag caaagcagga acagcggcag aaaattgtgc aggccgaggg tgaggccgag
     3061 gctgccaaga tgcttggaga agcactgagc aagaaccctg gctacatcaa acttcgcaag
     3121 attcgagcag cccagaatat ctccaagacg atcgccacat cacagaatcg tatctatctc
     3181 acagctgaca accttgtgct gaacctacag gatgaaagtt tcaccagggg aagtgacagc
     3241 ctcatcaagg gtaagaaaaa gcttcgaatt ctgcagtcga cggtaccgcg ggcccgggat
     3301 ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     3361 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     3421 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     3481 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     3541 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     3601 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     3661 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     3721 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     3781 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     3841 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     3901 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     3961 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     4021 gagctgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     4081 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     4141 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4201 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4261 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     4321 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     4381 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     4441 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     4501 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     4561 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     4621 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     4681 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     4741 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4801 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4861 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     4921 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4981 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5041 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     5101 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     5161 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     5221 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     5281 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     5341 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     5401 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     5461 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     5521 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     5581 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcg
//
