LOCUS       sequence-489300-        9268 bp    DNA     circular SYN 15-OCT-2025
DEFINITION  A piggyBac vector containing the COX5B 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 9268)
  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 9268)
  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: pJL347
FEATURES             Location/Qualifiers
     source          1..9268
                     /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   6722..6756
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7247..7263)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7247..7263)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7260..7282)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7271..7287
                     /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(7295..7301,7302..7319,7320..7325))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7340..7361
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7478..7495)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7649..8237)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7729..7748)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8408..9199,9200..9268))
                     /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     9031..9050
                     /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 acgcgtcaaa ggagacctga gggactgggt gtgagttgac agtggtggga
     3181 gctggaagca gcaagggatg cggggacatc tggggagcct ggacctgacc ttggagctca
     3241 ctcacacaat ctaaataatc ccagagtgag gctgaggttt tcctgagttg gatgagggcc
     3301 tgcgaaaatt tcctattatt aggcaagcct actatgagag tgagaagcct aaactcaatt
     3361 tccattttat agcctggagt aaaccacttc tggtcaggca gcagggaatc ataacatatc
     3421 agagctgaat acctcactgg tttgctgagg ggcaagttag taggtgttcc tcagaaagga
     3481 ttttgtggtc aattaagaca gggaaactct ggcttaaact gcaggtaaat taaataagag
     3541 gtctttactg caagcctaat cagagccttt attatgctag tgtccatctc caaaagggtg
     3601 gtcatcaaat gcaggccctc cctgtcttga ccaagtcctt tttcttgaga tcatgtcttg
     3661 agacttggat gccatgcaat acactttggg aaatactaac ctagttgaac gacagtttta
     3721 ctgatgtaga aatggcggcc caaaggataa aaagtagctg aactagctgg tggggctact
     3781 ttccagacca atgctttttt ctcctgccta agaccttaac cacagttgca aattaactac
     3841 agatcctcag aattttcgcc caattcaact tggaggggtt gcagtggagg tgagggagaa
     3901 gggagtaaca gtgaagccat tcaaagcaaa aaccaccttt aaagaaattt tggccttaga
     3961 atacagggtt tttttttttt ttttttaatt ggtggagggg gcacccagga agaccagacg
     4021 tgctcctgtg aagcaacgct ccaccacgtc cttttgaaaa taccgtagcc tgcatgagac
     4081 aggcgggagc acctaagcct tcaactctcc ccatgttgcc tccaattgac ctcactgggc
     4141 caggaaaagc cctataaact ggttttgtat actctatcca agtagtaaat ttactaaaat
     4201 tactggatcc tactatatac cagacctctg caaggcattg aagattcaat gatgagcatt
     4261 aattcccact ctaaggggat ttttctatct ggtagggaaa ctcagatacc gcatgcagct
     4321 agcctgtcac agggctatac tgactaaagc tcaccctaag aaatttgcac atcttagcct
     4381 atgggcaacg ggaagccatg gacggaatat gttttccgtt tcaaacaagg aactccagtc
     4441 acagcaggct ctaactccat ttcctattat caactcccct tcctgctcct cctcccttag
     4501 tagcctcgaa aaagagtggc cacttccttc acatcaccag ggccttggtc ctggtagggg
     4561 gtcccgcccc cggccctgtc ctcctcccca ggccgggacc tccgcgcctc ccacacacag
     4621 tacaactcta acacgaaact gggccaggac tgcaggtagg tcccgtccaa tgcgacacgc
     4681 tcgcgacggc ccacgcactg ccagaaccag ccctgccact aaccagccta gtaaggaacg
     4741 cagtgatctt taggagccca cttctaaatt caaatcattt ggggttcagt aagtctcttc
     4801 acagacgggg acctaatagc ttccattagc agcctccacg tttgctgaac acgactacgg
     4861 ggccagaccc acaggaactc caatgtggtt cagccgctca actccaggga ccaaaaagac
     4921 tttcccacct cccagcgtta ttaaaggcaa ccccgccccc aagcctcagc cactacgcgg
     4981 tgcagaaaga ggcagagctg ggcatgcgca atttccgccc ccgccacagt cttgggagtg
     5041 ggcgggctcg agggctggga ggaagccgaa agctgaccga gaggagaaag aagcccgccc
     5101 ccggaagtcc ctcctgtctc tgcagcttgt tcccggaagt tttgctgcta gtcgcggacg
     5161 caatggcttc aaggttactt cgcggagctg gaacgctggc cgcgcaggcc ctgagggctc
     5221 gcggccccag tggcgcggct gcgatgcgct ccatggcatc tggaggtggt gttcccactg
     5281 atgaagagca ggcgactggg ttggagaggg agatcatgct ggctgcaaag aagggactgg
     5341 acccatacaa tgtactggcc ccaaagggag cttcaggcac cagggaagac cctaatttag
     5401 tccccagcat cagcaacaag agaatagtag gctgcatctg tgaagaggac aataccagcg
     5461 tcgtctggtt ttggctgcac aaaggcgagg cccagcgatg cccccgctgt ggagcccatt
     5521 acaagctggt gccccagcag ctggcacact gagcacctgc actaaattac tcaaaatgtg
     5581 ctgtaaagtt tcttctttcc agtaaagact agccattgca ttggctcctt ctcccataga
     5641 tggctggtct tatttcttac ccgtattctt tggtaggcat ggaatatgct tattttggga
     5701 aaagctgtct gttaatgcta gcttgccatc cacttactga aagtgtataa ccagtgtata
     5761 gtgcttagat taataataag aatagatcga caacccgtaa tgcaatgaat gggaccacct
     5821 ggtatgagag aaaggggcgg gctgaggagt caggctgaca ggacttaaaa tattggctcc
     5881 atcatttggc tctatctctg tggacatgtt ctctgggggt caagtacaac tacaaaaagg
     5941 taagtacctt acaaggtgct taatatgtaa agagctgagt ggatagtagg tcctcacagt
     6001 atcctcatag agacagggtg gccttgtgaa gagagttttg gggattttga agtcttgagg
     6061 gacccaggtg caaagtaaga attcatgggg ataccagcag caaaattcag aatatggaac
     6121 aatttcagaa atcactgacc atgtttcttc aaataaattg caaggagtgg ggggaaaagc
     6181 tagggagaac atgtggagaa acagataagg gagataccag tcagcagcat gcacaccttg
     6241 tttagatctt gatttgaaaa caattataaa ggaaaaaaaa ttgagaagaa tttgaactct
     6301 ggatatttga tatgtagaca tttaccaccc tttcaggtat ttatatagtt ttgttttgtt
     6361 ttttttaatg cttgagatac gcacaggaat aagggagggg agaaatgcca aaaaagccca
     6421 ctttagctgc ttcatacctg tgacaccttg ggacaagtta aaatctcaaa accctcttac
     6481 ctacaaaatg gaaatacctt ttcattttgt gtggagatta gccagaggtg gcttcagtta
     6541 ctatcttcac aaaattggtc gaaagtaggt atttcagtgg gagctgatag agtcagagaa
     6601 aagccccttc ccctcatctt gcccatcctg aagggtttaa atcaagcggc gactgagatg
     6661 tcctaaatgc acagcgacgg attcgcgcta tttagaaaga gagagcaata tttcaagaat
     6721 gcatgcgtca attttacgca gactatcttt ctagggttaa tctagctgca tcaggatcat
     6781 atcgtcgggt cttttttccg gctcagtcat cgcccaagct ggcgctatct gggcatcggg
     6841 gaggaagaag cccgtgcctt ttcccgcgag gttgaagcgg catggaaaga gtttgccgag
     6901 gatgactgct gctgcattga cgttgagcga aaacgcacgt ttaccatgat gattcgggaa
     6961 ggtgtggcca tgcacgcctt taacggtgaa ctgttcgttc aggccacctg ggataccagt
     7021 tcgtcgcggc ttttccggac acagttccgg atggtcagcc cgaagcgcat cagcaacccg
     7081 aacaataccg gcgacagccg gaactgccgt gccggtgtgc agattaatga cagcggtgcg
     7141 gcgctgggat attacgtcag cgaggacggg tatcctggct ggatgccgca gaaatggaca
     7201 tggatacccc gtgagttacc cggcgggcgc gcttggcgta atcatggtca tagctgtttc
     7261 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     7321 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     7381 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     7441 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     7501 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     7561 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     7621 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     7681 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     7741 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     7801 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     7861 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     7921 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7981 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     8041 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     8101 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     8161 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     8221 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     8281 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     8341 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     8401 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     8461 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     8521 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     8581 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     8641 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     8701 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     8761 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     8821 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     8881 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     8941 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     9001 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     9061 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     9121 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     9181 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     9241 gggcgacacg gaaatgttga atactcat
//
