LOCUS       sequence-489276-        8250 bp    DNA     circular SYN 14-OCT-2025
DEFINITION  A piggyBac vector containing the COX7A2 coding sequence (CDS) 
            flanked by the 5' and 3' UTRs of the ACTB gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8250)
  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 8250)
  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: pJL359
FEATURES             Location/Qualifiers
     source          1..8250
                     /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   5704..5738
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6229..6245)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6229..6245)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6242..6264)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6253..6269
                     /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(6277..6283,6284..6301,6302..6307))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6322..6343
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6460..6477)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6631..7219)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6711..6730)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7390..8181,8182..8250))
                     /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     8013..8032
                     /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 tttttttgat atgaagtctc actctgtcgc ccaggctgga gtgcagtgaa
     3541 gcgatcttgg ctcactgcaa gctccacctt ctgggttcac gccattctcc tgcctcaacc
     3601 tcctgagtag ctgggactac aggcgcccgc caccacgccc gccttatttt ttgtattttt
     3661 agtagagaca gggtttcacg gctttagcca agatggtctg gatctcctga cctcatgatc
     3721 cgcccgcctc ggcttcccaa agtactggga ttacaggcgt gagccactgc gcctggcctc
     3781 atggctcttt ttggacaggg tcataagctt cttagatctg tcacccaatg taattaattg
     3841 atgtattggt taatactgtc tttagcagtg ctaggggatt ggtaagaaat ccacatcatc
     3901 tctgcgcttg ggaaatttac aatctagagt tggctaagtg gagaaaacat aagagagcac
     3961 atacaactaa caaattcaga ttatggacaa tgtacttaaa aaacccccac gccctagcta
     4021 ttggcacgtg gtaattttgc ccagagtttt ggttatatgc ataatccaga ggggtgtaga
     4081 aaaccaggtc ctttcagtac cgatctaagg aagacatgaa ttggggtgtt tataaacctt
     4141 ggtgaggaag agaattttaa ctgggactgg taacttggtg ggcccattga catacagtag
     4201 attgtgaaga agaaaacccc aaggattata aggaatagtt ttaatattac taatttttat
     4261 cctttgctga aactatgggt ttcttatttg aaatggcaat caaataattt gtaagagttt
     4321 gtagaaagtg gaaagattat tgtaatacag ggtagatatt tgcatttgta catacttgag
     4381 ggtaagtaca agaaataaac acttgtaatg gggattactt aagcaaaatg gatgacaccg
     4441 acagcctttc tgtacaacaa ataattgctg tgcttctcta ctccagtaat gaatgtagtg
     4501 ccctgctgga tctgggacag caagactttt gagtccagat tcgccatccc atttcttgtt
     4561 ccctaacgtt tcctcagaag cgagcgagcc tcttccacaa tttggggaca ctgtggaaga
     4621 acttcttgct tggggagttc tgcagcagtc gaaggtacaa gaatataaaa actggtatct
     4681 cgacacagct cttacagcat attctcaaca caaggaacag taatgtgccg cagcaattcc
     4741 ttcggtggaa aaaactgact gtatccgcgt aagaggagta attctttttt gaattttatg
     4801 aggcttcaga gctgtatttc ggccaaccca gtctttgttc agtttctagc ggcttcaagg
     4861 ttgcccagtt ttacgccttc tcgctcaaaa cagacactag ttttctttga tagcctggtc
     4921 tcacattgac ttcatctaag cttattacat tcgtctcata aatcgtttgc tttttcccca
     4981 gccttagctc cagggaccag cctactgaac atcgaaaagg tcaggaacga gatcacggga
     5041 ctgagacggc gagccggcta gagcggatag agccgtactg ccgctctagt ttttcttttg
     5101 ctctctatgg ttaggagcgc aagcctctct tcggcccgga aagatttaag ttcgtgaatg
     5161 catacgcaag actcggaggt agttccaccg ccgagaccgc gtccgccccg cgagcacaga
     5221 gcctcgcctt tgccgatccg ccgcccgtcc acacccgccg ccagctcacc atgctgcgga
     5281 atctgctggc tcttcgtcag attgggcaga ggacgataag aacagcaagc cgcaggcatt
     5341 ttaaaaataa agttccggag aagcaaaaac tgttccagga ggatgatgaa attccactgt
     5401 atctaaaggg tggggtagct gatgccctcc tgtatagagc caccatgatt cttacagttg
     5461 gtggaacagc atatgccata tatgagctgg ctgtggcttc atttcccaag aagcaggagt
     5521 gacttttggt ctccctggga gtgggtggag gcagccaggg cttacctgta cactgacttg
     5581 agaccagttg aataaaagtg cacaccttaa aaatgagttt aaaccaagcg gtgactgaga
     5641 tgtcctaaat gcacagcgac ggattcgcgc tatttagaaa gagagagcaa tatttcaaga
     5701 atgcatgcgt caattttacg cagactatct ttctagggtt aatctagctg catcaggatc
     5761 atatcgtcgg gtcttttttc cggctcagtc atcgcccaag ctggcgctat ctgggcatcg
     5821 gggaggaaga agcccgtgcc ttttcccgcg aggttgaagc ggcatggaaa gagtttgccg
     5881 aggatgactg ctgctgcatt gacgttgagc gaaaacgcac gtttaccatg atgattcggg
     5941 aaggtgtggc catgcacgcc tttaacggtg aactgttcgt tcaggccacc tgggatacca
     6001 gttcgtcgcg gcttttccgg acacagttcc ggatggtcag cccgaagcgc atcagcaacc
     6061 cgaacaatac cggcgacagc cggaactgcc gtgccggtgt gcagattaat gacagcggtg
     6121 cggcgctggg atattacgtc agcgaggacg ggtatcctgg ctggatgccg cagaaatgga
     6181 catggatacc ccgtgagtta cccggcgggc gcgcttggcg taatcatggt catagctgtt
     6241 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6301 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6361 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6421 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6481 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6541 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6601 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6661 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6721 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6781 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6841 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6901 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6961 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7021 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     7081 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7141 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7201 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7261 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7321 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7381 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7441 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7501 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7561 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7621 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7681 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7741 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7801 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7861 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7921 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7981 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     8041 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     8101 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     8161 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     8221 aagggcgaca cggaaatgtt gaatactcat
//
