LOCUS       sequence-489271-        9361 bp    DNA     circular SYN 14-OCT-2025
DEFINITION  A piggyBac vector containing the NDUFB9 coding sequences, modified 
            with silent mutations to allow for PCR-based analysis, along with 
            endogenous promoter and UTRs..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9361)
  AUTHORS   Luo J, Khandwala S, Hu J, Lee SY, Hickey KL, Levine ZG, Harper JW, 
            Ting AY, Weissman JS
  TITLE     Proximity-specific ribosome profiling reveals the logic of localized
            mitochondrial translation.
  JOURNAL   Cell. 2025 Aug 25:S0092-8674(25)00916-X. doi: 
            10.1016/j.cell.2025.08.002.
  PUBMED    40876456
REFERENCE   2  (bases 1 to 9361)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2025 
            Aug 25:S0092-8674(25)00916-X. doi: 10.1016/j.cell.2025.08.002."
COMMENT     Sequence Label: pJL257
FEATURES             Location/Qualifiers
     source          1..9361
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(268..289)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     480..499
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     713..735
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     727..744
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   874..936
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     misc_feature    1403..1991
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1509..1520
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1918..1938
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2016..2618)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGAS"
     primer_bind     complement(2102..2122)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2599..2618
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(2630..3130)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2693..2711)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     repeat_region   6815..6849
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7340..7356)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7340..7356)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7353..7375)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7364..7380
                     /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(7388..7394,7395..7412,7413..7418))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7433..7454
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7571..7588)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7742..8330)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7822..7841)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8501..9292,9293..9361))
                     /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     9124..9143
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgt gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaac aaaaactcaa
     1081 aatttcttct ataaagtaac aaaactttta tgagggacag ccccccccca aagcccccag
     1141 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     1201 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     1261 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     1321 cacctggggg gatacgggga aaaggcctcc agggtttatt acagggacag cagagatcca
     1381 gtttggttaa ttagctagcg atgcggggag gcggcccaaa gggagatccg actcgtctga
     1441 gggcgaaggc gaagacgcgg aagaggccgc agagccggca gcaggccgcg ggaaggaagg
     1501 tccgctggat tgagggccga agggacgtag cagaaggacg tcccgcgcag aatccaggtg
     1561 gcaacacagg cgagcagcca aggaaaggac gatgatttcc ccgacaacac cacggaattg
     1621 tcagtgccca acagccgagc ccctgtccag cagcgggcaa ggcaggcggc gatgagttcc
     1681 gccgtggcaa tagggagggg gaaagcgaaa gtcccggaaa ggagctgaca ggtggtggca
     1741 atgccccaac cagtgggggt tgcgtcagca aacacagtgc acaccacgcc acgttgcctg
     1801 acaacgggcc acaactcctc ataaagagac agcaaccagg atttatacaa ggaggagaaa
     1861 atgaaagcca tacgggaagc aatagcatga tacaaaggca ttaaagcagc gtatccacat
     1921 agcgtaaaag gagcaacata gttaagaata ccagtcaatc tttcacaaat tttgtaatcc
     1981 agaggttgat taccgataag cttgatatcg aattcttaac tggcaccggg cttgcgggtc
     2041 atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg cggtgacggt gaagccgagc
     2101 cgctcgtaga aggggaggtt gcggggcgcg gaggtctcca ggaaggcggg caccccggcg
     2161 cgctcggccg cctccactcc ggggagcacg acggcgctgc ccagaccctt gccctggtgg
     2221 tcgggcgaga cgccgacggt ggccaggaac cacgcgggct ccttgggccg gtgcggcgcc
     2281 aggaggcctt ccatctgttg ctgcgcggcc agccgggaac cgctcaactc ggccatgcgc
     2341 gggccgatct cggcgaacac cgcccccgct tcgacgctct ccggcgtggt ccagaccgcc
     2401 acggcggcgc cgtcgtccgc gacccacacc ttgccgatgt cgagcccgac gcgcgtgagg
     2461 aagagttctt gcagctcggt gacccgctcg atgtggcggt ccggatcgac ggtgtggcgc
     2521 gtggcggggt agtcggcgaa cgcggcggcg agggtgcgta cggccctggg gacgtcgtcg
     2581 cgggtggcga ggcgcaccgt gggcttgtac tcggtcatgg tggtaccccc tggggagaga
     2641 ggtcggtgat tcggtcaacg agggagccga ctgccgacgt gcgctccgga ggcttgcaga
     2701 atgcggaaca ccgcgcgggc aggaacaggg cccacactac cgccccacac cccgcctccc
     2761 gcaccgcccc ttcccggccg ctgctctcgg cgcgccctgc tgagcagccg ctattggcca
     2821 cagcccatcg cggtcggcgc gctgccattg ctccctggcg ctgtccgtct gcgagggtac
     2881 tagtgagacg tgcggcttcc gtttgtcacg tccggcacgc cgcgaaccgc aaggaacctt
     2941 cccgacttag gggcggagca ggaagcgtcg ccggggggcc cacaagggta gcggcgaaga
     3001 tccgggtgac gctgcgaacg gacgtgaaga atgtgcgaga cccagggtcg gcgccgctgc
     3061 gtttcccgga accacgccca gagcagccgc gtccctgcgc aaacccaggg ctgccttgga
     3121 aaaggcgcaa acgcgtccca aacagtcatt gtactacgcc cctaggaaaa aaatataaat
     3181 accataccag tatctgcctt aagagtttga tctagtggga aaggcatact catatttaat
     3241 tataaataat gtggtataag ttttaaggag tgagtaggca gacaggcaac agggtagttg
     3301 gtaggcagag gaatattttg gccccaaact gccactctaa aaccttgtca gaggtaacaa
     3361 acagcatgca caagtgttta tagagcagcc cagggatggg ctagaaaagt gggcaggact
     3421 ggaccacaga agggctttgt atggtgtgta agcatttagg ctatcctgag gagcgtaaat
     3481 gaaagatttg aagcaaaaga aacatttagc tgatttgatt tttagataca ccaaaatgga
     3541 gtctacatgg aggatggatt taagacatta acttgaggca ggaatttggt tgtataacat
     3601 aaacacactt ttaaaaagtt atgtggcctt tctttacgga gtacttcatg gctctttatt
     3661 ccatctccct catcaattat atcaaaggtt aggatggaaa ggctaagaca cggcaatgtc
     3721 ttatttgaca acccctcccc aaacccctac tccaggcttt cccacagcaa tttcaaaacc
     3781 tttttcctgg cacttaacat attctcccct gcatttgctt ttgactgtac agtacatagg
     3841 tcttaggtct ccagtctgaa aactgcacca ctagatttta cagttccaca ggacttagca
     3901 caacgtcttt gaaaaaaaaa aatactgctt aaataactct cagatctaaa tgtataatcc
     3961 taatctcttg attgccagac cacattccca attgcccaga tatctctacc tggatgtcct
     4021 ataattatct caaacttaac ctaatccaaa tggacctggg tacctctctt caccgccaaa
     4081 ctagtcatgc ttccatttca tatttaagta cttttccaaa ccctgagtca tctttgttca
     4141 ctctctacta ccggccatga acactcagct gctaaatgct gcacacatct cttccatttc
     4201 tctctctttt ccattaccct agtttaagct cttgaaatgc ttccctacct ctccctattt
     4261 cccacaaaac ctacataata gctatcagga ttaccttgct aaagcagagg tgtgatcact
     4321 gtcatttccc accttcaggt agctggaaac cccacaaaac gaggatcatg cccatttaaa
     4381 tcccgcagca ttctccctct cgttctactg atgccgactg gatcattgta cttgccaagt
     4441 acagtttgtt tttctcccca ttacttgtgt tttctagttt ctccaccaga tggtgatgac
     4501 agtacaactg ggagctttgt gccctgtgtt cctaggccag tgccaggccc acttgaggcg
     4561 ctcactaact atgatatgaa cgaatgaatg cttgagtgaa ttggtcccca gaagcggtca
     4621 gcctctccgc caaggcagag gggaagggcc gggaggtgca cgtgggatga atggaaacat
     4681 agtgattggc ctttttggag ttttcttctt aaaaaactga atacattaca gatgttgaaa
     4741 acagcacaaa aatcataact gtacggcttg ttaattttta caaataaata caccctcgtg
     4801 aatccagaac ctagatcaag aaccagagca ttaccagcac ccagaaacct ccccgcgccc
     4861 tcctccactg agcggccctt tcaaccctta caatgccgga gggcgcaatt cctgggtcct
     4921 gtgaccttgg tgaccgacgc ccgggacaag tcagaaagac ccaagggcgt ggaaaacgct
     4981 cctcttaccg ataaacttga agcgactcat gactgcgcat ggaggacctc cccagcggaa
     5041 gcggaagtgg ccgccggcaa ctccgccctt ccggctggcc ccgctcagtc acccgcagca
     5101 ggcgtgcagt ttcccggctc tccgcgcggc cggggaaggt cagcgccgta atggcgttct
     5161 tggcgtcggg accctacctg acccatcagc aaaaggtgtt gcggctttat aagcgggcgc
     5221 tacgccacct cgagtcgtgg tgcgtccaga gagacaaata ccgatacttt gcttgtttga
     5281 tgagagcccg gtttgaagaa cataagaatg aaaaggatat ggcgaaggcc acccagctgc
     5341 tgaaggaggc cgaggaagaa ttctggtacc gtcagcatcc acagccatac atcttccctg
     5401 actctcctgg gggcaccagc tatgagcggt acgattgcta caaggtccca gaatggtgct
     5461 tagatgactg gcatccttct gagaaggcaa tgtatcctga ttactttgcc aagagagaac
     5521 agtggaagaa actgcggagg gaaagctggg aacgagaggt taagcagctg caggaggaaa
     5581 cgccacctgg tggtccttta actgaagctt tgccccctgc ccgaaaggaa ggtgatttgc
     5641 ccccactgtg gtggtatatt gtgaccagac cccgggagcg gcccatgtag aaagagagag
     5701 acctcatctt tcatgcttgc aagtgaaata tgttacagaa catgcacttg ccctaataaa
     5761 aaatcagtga aatggtctct ggtatgactg acgttcttct aagaattact gtgatgccaa
     5821 gtgactgtgc cactgtcatt gttttatcaa tcgtgcatgt aaagaaaatg acaggacatg
     5881 tagtcagtca gctgtgttct ctgcaataaa tcactgatag catatgttac cagtaaggaa
     5941 aaactgaggc agaaccagat tgaggcgaac aaactctcaa gtcaaaggcc taattagctg
     6001 tggtaactag aaggaataaa caaagccaag aacagaaata ctgaggattt aaaaataacc
     6061 tttttggttt tctaaagatt tcactaatgg aacttagttt ttattagaag aacaactctc
     6121 cctaagaagt acatagtcaa caaataagca tggaatttgt ggagttctgt caggggtgga
     6181 ggtttgatgg ttggggttaa atggtccaat ctgggataaa actttgtggc gcttatttta
     6241 gaagcagttt tccagctttg cttctgctac cgtagcttgg attgtctgcg gactcaaagg
     6301 taaaggaata acagaactcc ttaaatgttt cagaagcttt gtttttgttt tttaaagata
     6361 aagcagtttt taaaactttc aacccaaaac agctgctttt agttcttgat ccctcattag
     6421 ctagaactgt tttcaaatag tttcaggcaa ctaatgatct tatagccatc acaaaaatat
     6481 tttgtcactt ttctacgtaa gctcaatttg agtcccagtc catcttctaa catttgacta
     6541 agacctcaaa ctgaattgta acaagacttg aaatacagga cttttttaat ctagtggttc
     6601 tttatttata acgtaggctg aagcaattgt tacaatgtag gagggagaca cattacacaa
     6661 ttgttattta tacaagttta gaacaaaaaa ctgcacaggg agaggtcaac tctcagtaca
     6721 aactagcgtt taaaccaagc ggcgactgag atgtcctaaa tgcacagcga cggattcgcg
     6781 ctatttagaa agagagagca atatttcaag aatgcatgcg tcaattttac gcagactatc
     6841 tttctagggt taatctagct gcatcaggat catatcgtcg ggtctttttt ccggctcagt
     6901 catcgcccaa gctggcgcta tctgggcatc ggggaggaag aagcccgtgc cttttcccgc
     6961 gaggttgaag cggcatggaa agagtttgcc gaggatgact gctgctgcat tgacgttgag
     7021 cgaaaacgca cgtttaccat gatgattcgg gaaggtgtgg ccatgcacgc ctttaacggt
     7081 gaactgttcg ttcaggccac ctgggatacc agttcgtcgc ggcttttccg gacacagttc
     7141 cggatggtca gcccgaagcg catcagcaac ccgaacaata ccggcgacag ccggaactgc
     7201 cgtgccggtg tgcagattaa tgacagcggt gcggcgctgg gatattacgt cagcgaggac
     7261 gggtatcctg gctggatgcc gcagaaatgg acatggatac cccgtgagtt acccggcggg
     7321 cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     7381 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     7441 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     7501 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     7561 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     7621 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     7681 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     7741 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     7801 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     7861 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7921 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7981 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     8041 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     8101 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     8161 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     8221 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     8281 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     8341 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     8401 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     8461 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     8521 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     8581 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     8641 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     8701 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     8761 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     8821 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     8881 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     8941 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     9001 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     9061 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     9121 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     9181 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     9241 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     9301 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     9361 t
//
