LOCUS       sequence-489308-        8426 bp    DNA     circular SYN 15-OCT-2025
DEFINITION  A piggyBac vector containing the NDUFB9 coding sequence under the 
            control of its endogenous promoter and 5' UTR, followed by the ACTB 
            3' UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8426)
  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 8426)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 15, 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: pJL308
FEATURES             Location/Qualifiers
     source          1..8426
                     /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   5880..5914
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6405..6421)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6405..6421)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6418..6440)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6429..6445
                     /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(6453..6459,6460..6477,6478..6483))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6498..6519
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6636..6653)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6807..7395)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6887..6906)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7566..8357,8358..8426))
                     /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     8189..8208
                     /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 ttaattaact
     5701 tttggtctcc ctgggagtgg gtggaggcag ccagggctta cctgtacact gacttgagac
     5761 cagttgaata aaagtgcaca ccttaaaaat gagtttaaac caagcggcga ctgagatgtc
     5821 ctaaatgcac agcgacggat tcgcgctatt tagaaagaga gagcaatatt tcaagaatgc
     5881 atgcgtcaat tttacgcaga ctatctttct agggttaatc tagctgcatc aggatcatat
     5941 cgtcgggtct tttttccggc tcagtcatcg cccaagctgg cgctatctgg gcatcgggga
     6001 ggaagaagcc cgtgcctttt cccgcgaggt tgaagcggca tggaaagagt ttgccgagga
     6061 tgactgctgc tgcattgacg ttgagcgaaa acgcacgttt accatgatga ttcgggaagg
     6121 tgtggccatg cacgccttta acggtgaact gttcgttcag gccacctggg ataccagttc
     6181 gtcgcggctt ttccggacac agttccggat ggtcagcccg aagcgcatca gcaacccgaa
     6241 caataccggc gacagccgga actgccgtgc cggtgtgcag attaatgaca gcggtgcggc
     6301 gctgggatat tacgtcagcg aggacgggta tcctggctgg atgccgcaga aatggacatg
     6361 gataccccgt gagttacccg gcgggcgcgc ttggcgtaat catggtcata gctgtttcct
     6421 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     6481 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6541 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6601 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6661 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6721 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6781 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6841 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6901 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6961 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     7021 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     7081 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     7141 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     7201 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     7261 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7321 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7381 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7441 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     7501 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7561 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7621 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7681 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7741 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7801 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7861 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7921 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7981 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     8041 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     8101 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     8161 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     8221 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     8281 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     8341 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     8401 gcgacacgga aatgttgaat actcat
//
