LOCUS       sequence-199361-        5638 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mitochondrial marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5638)
  AUTHORS   Kwon SK, Sando R 3rd, Lewis TL, Hirabayashi Y, Maximov A, Polleux F
  TITLE     LKB1 Regulates Mitochondria-Dependent Presynaptic Calcium Clearance 
            and Neurotransmitter Release Properties at Excitatory Synapses along
            Cortical Axons.
  JOURNAL   PLoS Biol. 2016 Jul 18;14(7):e1002516. doi: 
            10.1371/journal.pbio.1002516. eCollection 2016 Jul.
  PUBMED    27429220
REFERENCE   2  (bases 1 to 5638)
  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: "PLoS 
            Biol."; date: "2016-07-18"; volume: "14(7):e1002516. doi"; pages: " 
            10.1371/journal.pbio.1002516. eCollection 2016 Jul"
COMMENT     Sequence Label: pCAG mito-mTagBFP2
FEATURES             Location/Qualifiers
     source          1..5638
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(42..629)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(122..141)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(800..1591,1592..1660))
                     /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     1423..1442
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1661..1765)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        1796..2175
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2177..2454
                     /label=chicken beta-actin promoter
     intron          2455..3467
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3391..3413
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3475..3494
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3528..3602
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             3630..4331
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(4422..4441)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4487..4542
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4541..4560)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4933..4949)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4933..4949)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4946..4968)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4957..4973
                     /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(4981..4987,4988..5005,5006..5011))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5026..5047
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5106..5301
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5152..5287
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5307..5441
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5344..5363)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5398..5417
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5509..5526)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
       61 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      121 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      181 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      241 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      301 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      361 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      421 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      481 agacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
      541 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
      601 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
      661 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
      721 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
      781 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
      841 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
      901 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
      961 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     1021 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     1081 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     1141 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     1201 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     1261 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     1321 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     1381 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     1441 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     1501 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     1561 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     1621 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     1681 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     1741 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gggtcgacat
     1801 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     1861 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     1921 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     1981 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     2041 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     2101 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     2161 atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc
     2221 ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg
     2281 gcgggggggg ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg
     2341 gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt
     2401 atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc
     2461 gctgcgttgc cttcgccccg tgccccgctc cgcgccgcct cgcgccgccc gccccggctc
     2521 tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt
     2581 aattagcgct tggtttaatg acggctcgtt tcttttctgt ggctgcgtga aagccttaaa
     2641 gggctccggg agggcccttt gtgcgggggg gagcggctcg gggggtgcgt gcgtgtgtgt
     2701 gtgcgtgggg agcgccgcgt gcggcccgcg ctgcccggcg gctgtgagcg ctgcgggcgc
     2761 ggcgcggggc tttgtgcgct ccgcgtgtgc gcgaggggag cgcggccggg ggcggtgccc
     2821 cgcggtgcgg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg
     2881 gtgagcaggg ggtgtgggcg cggcggtcgg gctgtaaccc ccccctgcac ccccctcccc
     2941 gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc
     3001 tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc
     3061 gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag
     3121 gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc
     3181 ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg
     3241 ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg
     3301 cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg
     3361 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
     3421 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
     3481 tgctggttat tgtgctgtct catcattttg gcaaagaatt gctcgagatg tccgtcctga
     3541 cgccgctgct gctgcggggc ttgacaggct cggcccggcg gctcccagtg ccgcgcgcca
     3601 agatccattc gttgggggat ccaccggtca tgagcgagct gattaaggag aacatgcaca
     3661 tgaagctgta catggagggc accgtggaca accatcactt caagtgcaca tccgagggcg
     3721 aaggcaagcc ctacgagggc acccagacca tgagaatcaa ggtggtcgag ggcggccctc
     3781 tccccttcgc cttcgacatc ctggctacta gcttcctcta cggcagcaag accttcatca
     3841 accacaccca gggcatcccc gacttcttca agcagtcctt ccctgagggc ttcacatggg
     3901 agagagtcac cacatacgaa gacgggggcg tgctgaccgc tacccaggac accagcctcc
     3961 aggacggctg cctcatctac aacgtcaaga tcagaggggt gaacttcaca tccaacggcc
     4021 ctgtgatgca gaagaaaaca ctcggctggg aggccttcac cgagacgctg taccccgctg
     4081 acggcggcct ggaaggcaga aacgacatgg ccctgaagct cgtgggcggg agccatctga
     4141 tcgcaaacgc caagaccaca tatagatcca agaaacccgc taagaacctc aagatgcctg
     4201 gcgtctacta tgtggactac agactggaaa gaatcaagga ggccaacaac gagacctacg
     4261 tcgagcagca cgaggtggca gtggccagat actgcgacct ccctagcaaa ctggggcaca
     4321 agcttaatta agcggccgca attcactcct caggtgcagg ctgcctatca gaaggtggtg
     4381 gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa
     4441 aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt
     4501 attttcattg caatagtgtg ttggaatttt ttgtgtctat cactcggaag gacatatggg
     4561 agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc
     4621 catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag
     4681 ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta gatttttttt
     4741 atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta
     4801 ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta
     4861 tggagatccc tcgacctgca ccgtcgacca gctggtcgac ggtgcaccgt cgaccagctt
     4921 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     4981 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5041 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg
     5101 gatccgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     5161 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     5221 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     5281 ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat
     5341 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5401 gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg
     5461 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5521 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5581 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccag
//
