LOCUS       sequence-489280-        9389 bp    DNA     circular SYN 14-OCT-2025
DEFINITION  A piggyBac vector containing the COX7A2 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 9389)
  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 9389)
  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: pJL357
FEATURES             Location/Qualifiers
     source          1..9389
                     /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   6843..6877
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7368..7384)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7368..7384)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7381..7403)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7392..7408
                     /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(7416..7422,7423..7440,7441..7446))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7461..7482
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7599..7616)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7770..8358)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7850..7869)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8529..9320,9321..9389))
                     /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     9152..9171
                     /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 acgcgttggt gagggacgga ttaactatgt taaatgctgc taatagatgg
     3181 agaaggtgaa aattgagatt gacagtttat ttagcaacct ggaagctatt ggtgattcta
     3241 aacaagagca gtaaaagcct gcccaaaatg ggatcaaaaa aagattagga ggaggagaag
     3301 gagagacaag tataaagaaa tctataggat ggtaacggta atcaaaggag tttgttcgtt
     3361 tgtttttaat ttgggaatgt gttaatagat ggaatgatcc agtaaagggg gaaattgtac
     3421 tgaataacat gcatacaaga atctatgtct actttaggat ctatttaatg tgaattcatg
     3481 gctttttttt tttttttttg atatgaagtc tcactctgtc gcccaggctg gagtgcagtg
     3541 aagcgatctt ggctcactgc aagctccacc ttctgggttc acgccattct cctgcctcaa
     3601 cctcctgagt agctgggact acaggcgccc gccaccacgc ccgccttatt ttttgtattt
     3661 ttagtagaga cagggtttca cggctttagc caagatggtc tggatctcct gacctcatga
     3721 tccgcccgcc tcggcttccc aaagtactgg gattacaggc gtgagccact gcgcctggcc
     3781 tcatggctct ttttggacag ggtcataagc ttcttagatc tgtcacccaa tgtaattaat
     3841 tgatgtattg gttaatactg tctttagcag tgctagggga ttggtaagaa atccacatca
     3901 tctctgcgct tgggaaattt acaatctaga gttggctaag tggagaaaac ataagagagc
     3961 acatacaact aacaaattca gattatggac aatgtactta aaaaaccccc acgccctagc
     4021 tattggcacg tggtaatttt gcccagagtt ttggttatat gcataatcca gaggggtgta
     4081 gaaaaccagg tcctttcagt accgatctaa ggaagacatg aattggggtg tttataaacc
     4141 ttggtgagga agagaatttt aactgggact ggtaacttgg tgggcccatt gacatacagt
     4201 agattgtgaa gaagaaaacc ccaaggatta taaggaatag ttttaatatt actaattttt
     4261 atcctttgct gaaactatgg gtttcttatt tgaaatggca atcaaataat ttgtaagagt
     4321 ttgtagaaag tggaaagatt attgtaatac agggtagata tttgcatttg tacatacttg
     4381 agggtaagta caagaaataa acacttgtaa tggggattac ttaagcaaaa tggatgacac
     4441 cgacagcctt tctgtacaac aaataattgc tgtgcttctc tactccagta atgaatgtag
     4501 tgccctgctg gatctgggac agcaagactt ttgagtccag attcgccatc ccatttcttg
     4561 ttccctaacg tttcctcaga agcgagcgag cctcttccac aatttgggga cactgtggaa
     4621 gaacttcttg cttggggagt tctgcagcag tcgaaggtac aagaatataa aaactggtat
     4681 ctcgacacag ctcttacagc atattctcaa cacaaggaac agtaatgtgc cgcagcaatt
     4741 ccttcggtgg aaaaaactga ctgtatccgc gtaagaggag taattctttt ttgaatttta
     4801 tgaggcttca gagctgtatt tcggccaacc cagtctttgt tcagtttcta gcggcttcaa
     4861 ggttgcccag ttttacgcct tctcgctcaa aacagacact agttttcttt gatagcctgg
     4921 tctcacattg acttcatcta agcttattac attcgtctca taaatcgttt gctttttccc
     4981 cagccttagc tccagggacc agcctactga acatcgaaaa ggtcaggaac gagatcacgg
     5041 gactgagacg gcgagccggc tagagcggat agagccgtac tgccgctcta gtttttcttt
     5101 tgctctctat ggttaggagc gcaagcctct cttcggcccg gaaagattta agttcgtgaa
     5161 tgcatacgca agactcggag gtagttccgg ttccggcgtg gccattttcg ttggtggtgt
     5221 tcagttgtgg cggttgctgg tcagtaacag ccaagatgct gcggaatctg ctggctcttc
     5281 gtcagattgg gcagaggacg ataagaacag caagccgcag gcattttaaa aataaagttc
     5341 cggagaagca aaaactgttc caggaggatg atgaaattcc actgtatcta aagggtgggg
     5401 tagctgatgc cctcctgtat agagccacca tgattcttac agttggtgga acagcatatg
     5461 ccatatatga gctggctgtg gcttcatttc ccaagaagca ggagtgactt cagtcatccc
     5521 agcaatcgct tggttcagtt tcattcagct ctctatggac cagtaatctg ataaataacc
     5581 gagctcttct ttggggatca atatttattg acttgtagta actgccacca ataaagcagt
     5641 ctttaccatg ctttgtctga ttttatcact ttatgccaat aatttcaact tttggtcagc
     5701 cttgtaaatc ttaggtaagg atcaaagaaa tgattctttt gccactaaac ttgtgaaagc
     5761 aacattattg tgatgttatt ttagtgaaat tttacttcta acagctcaaa attagtctca
     5821 acttctgtct caacttgccg tttgaatgca tattgctttt tccccctacc cactccagta
     5881 tatcttggaa tatgctagaa cccgggggta gtggaagcag attatctttg gctcctgggg
     5941 tctatatttc tttttttttt ttttttctca gctatgtgag agtatgctta aatattggat
     6001 atatcccaca gcttagaatt aggctttagt agaatggctt tgtaagatat ttcagtattt
     6061 gtttttttct acactccgag tgggttcagg taggtttctc tcatcaggaa ccaaccaaag
     6121 caggacagtt cctgggctcc ccagctatca tctaatggac ccacaactgt ttaacctaaa
     6181 cagtagttag gtgcttaata aatggtagcc ataataggct gctcataata ggaatgtttc
     6241 aaaaaggtct atttagggaa cacaaaatcc aaatagatac agagagtaag aatcaaaaag
     6301 aattacgatg ctgctctagg tagtttgctt cagggtttgc agaacccttc ctgctttgag
     6361 agaactatgc atgaagctgg aaatttcata gacttcaagt ttgggactcc cttttcctgg
     6421 gtcctgttgt gcatccatgt ggagagtagc atcttgtgaa ccacttcagg actttccttc
     6481 tatatatatc gagattggta attcgtcaag agatactgat tgcctactgt gtcctaactg
     6541 gttttaggta cctaggaatc actggtgaac aaaagcagat tcttcccctc atatgttata
     6601 gaaaggggtg gtaggaaaga gagtttaaaa ataaattgca cagtaggtta aaagataagt
     6661 tctatggaaa aagaccaaag gggagaactc acatgggtgt ggttggttgg ttgcagtatt
     6721 aaagttagga taggcttcaa caggaaggtc agattgttta aaccaagcgg cgactgagat
     6781 gtcctaaatg cacagcgacg gattcgcgct atttagaaag agagagcaat atttcaagaa
     6841 tgcatgcgtc aattttacgc agactatctt tctagggtta atctagctgc atcaggatca
     6901 tatcgtcggg tcttttttcc ggctcagtca tcgcccaagc tggcgctatc tgggcatcgg
     6961 ggaggaagaa gcccgtgcct tttcccgcga ggttgaagcg gcatggaaag agtttgccga
     7021 ggatgactgc tgctgcattg acgttgagcg aaaacgcacg tttaccatga tgattcggga
     7081 aggtgtggcc atgcacgcct ttaacggtga actgttcgtt caggccacct gggataccag
     7141 ttcgtcgcgg cttttccgga cacagttccg gatggtcagc ccgaagcgca tcagcaaccc
     7201 gaacaatacc ggcgacagcc ggaactgccg tgccggtgtg cagattaatg acagcggtgc
     7261 ggcgctggga tattacgtca gcgaggacgg gtatcctggc tggatgccgc agaaatggac
     7321 atggataccc cgtgagttac ccggcgggcg cgcttggcgt aatcatggtc atagctgttt
     7381 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     7441 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     7501 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     7561 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     7621 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     7681 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     7741 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     7801 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     7861 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     7921 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     7981 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     8041 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     8101 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     8161 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     8221 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     8281 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     8341 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     8401 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     8461 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     8521 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     8581 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     8641 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     8701 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     8761 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     8821 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     8881 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     8941 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     9001 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     9061 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     9121 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     9181 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     9241 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     9301 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     9361 agggcgacac ggaaatgttg aatactcat
//
