LOCUS       sequence-461302-        6818 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses APEX fused to PEX26 peroxisomal localization sequence in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6818)
  AUTHORS   Wyant GA, Jiang Q, Singh M, Qayyum S, Levrero C, Maron BA, Kaelin WG
            Jr
  TITLE     Induction of DEPP1 by HIF Mediates Multiple Hallmarks of Ischemic 
            Cardiomyopathy.
  JOURNAL   Circulation. 2024 Jun 17. doi: 10.1161/CIRCULATIONAHA.123.066628.
  PUBMED    38881449
REFERENCE   2  (bases 1 to 6818)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Circulation. 2024 Jun 17. doi: 10.1161/CIRCULATIONAHA.123.066628."
COMMENT     Sequence Label: pMXS-IP APEX-PEX26
FEATURES             Location/Qualifiers
     source          1..6818
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     27..47
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             301..770
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(900..917)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1071..1659)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1151..1170)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1823..2614,2615..2683))
                     /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     2446..2465
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2684..2788)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             2818..3411
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    3474..3831
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             3890..4306
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     4236..4258
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     4274..4290
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    4316..4690
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     4628..4644
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      4741..4750
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4750..4773
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4774..5520
                     /codon_start=1
                     /product="soybean ascorbate peroxidase, engineered to serve
                     as a monomeric tag for electron microscopy and live-cell 
                     proteomics (Lam et al., 2015)"
                     /label=APEX2
                     /note="more active than the original APEX due to the A134P 
                     mutation"
                     /translation="GKSYPTVSADYQDAVEKAKKKLRGFIAEKRCAPLMLRLAFHSAGT
                     FDKGTKTGGPFGTIKHPAELAHSANNGLDIAVRLLEPLKAEFPILSYADFYQLAGVVAV
                     EVTGGPKVPFHPGREDKPEPPPEGRLPDPTKGSDHLRDVFGKAMGLTDQDIVALSGGHT
                     IGAAHKERSGFEGPWTSNPLIFDNSYFTELLSGEKEGLLQLPSDKALLSDPVFRPLVDK
                     YAADEDAFFADYAEAHQKLSELGFADA"
     misc_feature    5766..6332
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5782..6334
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5926..5943)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6153..6172
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(6346..6818,1..130)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MVTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
ORIGIN
        1 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
       61 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
      121 cggtgcctga gtcgacgata aaataaaaga ttttatttag tctccagaaa aaggggggaa
      181 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
      241 ggaaaaatac ataactgaga atagagaagt tcagatcaag gtcaggaaca gatggaacag
      301 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      361 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
      421 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
      481 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
      541 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
      601 aaaagagccc acaacccctc actcggcgcg ccagtcctcc gattgactga gtcgcccggg
      661 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
      721 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca catgcagcat
      781 gtatcaaaat taatttggtt ttttttctta agtatttaca ttaaatggcc atagttgcat
      841 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
      901 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
      961 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     1021 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1081 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     1141 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     1201 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     1261 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     1321 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     1381 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     1441 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     1501 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     1561 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     1621 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     1681 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     1741 tcaaaaagga tcttcaccta gatccttttg cggccggccg caaatcaatc taaagtatat
     1801 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     1861 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     1921 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     1981 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     2041 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     2101 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     2161 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     2221 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     2281 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     2341 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     2401 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     2461 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     2521 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     2581 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     2641 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     2701 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     2761 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgcataat
     2821 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     2881 gaaaaataca taactgagaa tagaaaagtt cagatcaagg tcaggaacag atggaacagc
     2941 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     3001 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     3061 cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga
     3121 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
     3181 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
     3241 aaagagccca caacccctca ctcggcgcgc cagtcctccg attgactgag tcgcccgggt
     3301 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt
     3361 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcat ttgggggctc
     3421 gtccgggatc gggagacccc tgcccaggga ccaccgaccc accaccggga ggtaagctgg
     3481 ccagcaactt atctgtgtct gtccgattgt ctagtgtcta tgactgattt tatgcgcctg
     3541 cgtcggtact agttagctaa ctagctctgt atctggcgga cccgtggtgg aactgacgag
     3601 ttcggaacac ccggccgcaa ccctgggaga cgtcccaggg acttcggggg ccgtttttgt
     3661 ggcccgacct gagtccaaaa atcccgatcg ttttggactc tttggtgcac ccccctaata
     3721 ggagggatat gtggttctgg taggagacga gaacctaaaa cagttcccgc ctccgtctga
     3781 atttttgctt tcggtttggg accgaagccg cgccgcgcgt cttgtctgct gcagcatcgt
     3841 tctgtgttgt ctctgtctga ctgtgtttct gtatttgtct gaaaattagg gccagactgt
     3901 taccactccc ttaagtttga ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa
     3961 ccagtcggta gatgtcaaga agagacgttg ggttaccttc tgctctgcag aatggccaac
     4021 ctttaacgtc ggatggccgc gagacggcac ctttaaccga gacctcatca cccaggttaa
     4081 gatcaaggtc ttttcacctg gcccgcatgg acacccagac caggtcccct acatcgtgac
     4141 ctgggaagcc ttggcttttg acccccctcc ctgggtcaag ccctttgtac accctaagcc
     4201 tccgcctcct cttcctccat ccgccccgtc tctccccctt gaacctcctc gttcgacccc
     4261 gcctcgatcc tccctttatc cagccctcac tccttctcta ggcgccccca tatggccata
     4321 tgagatctta tatggggcac ccccgcccct tgtaaacttc cctgaccctg acatgacaag
     4381 agttactaac agcccctctc tccaagctca cttacaggct ctctacttag tccagcacga
     4441 agtctggaga cctctggcgg cagcctacca agaacaactg gaccgaccgg tggtacctca
     4501 cccttaccga gtcggcgaca cagtgtgggt ccgccgacac cagactaaga acctagaacc
     4561 tcgctggaaa ggaccttaca cagtcctgct gaccaccccc accgccctca aagtagacgg
     4621 catcgcagct tggatacacg ccgcccacgt gaaggctgcc gaccccgggg gtggaccatc
     4681 ctctagactg ccggatctag ctagttaatt aaggatccca gtgtggtggt acgggaattc
     4741 gccaccatgg actacaagga tgacgacgat aagggaaagt cttacccaac tgtgagtgct
     4801 gattaccagg acgccgttga gaaggcgaag aagaagctca gaggcttcat cgctgagaag
     4861 agatgcgctc ctctaatgct ccgtttggca ttccactctg ctggaacctt tgacaagggc
     4921 acgaagaccg gtggaccctt cggaactatc aagcaccctg ccgaactggc tcacagcgct
     4981 aacaacggtc ttgacatcgc tgttaggctt ttggagccac tcaaggcgga gttccctatt
     5041 ttgagctacg ccgatttcta ccagttggct ggcgttgttg ccgttgaggt cacgggtgga
     5101 cctaaggttc cattccaccc tggaagagag gacaagcctg agccaccacc agagggtcgc
     5161 ttgcccgatc ccactaaggg ttctgaccat ttgagagatg tgtttggcaa agctatgggg
     5221 cttactgacc aagatatcgt tgctctatct gggggtcaca ctattggagc tgcacacaag
     5281 gagcgttctg gatttgaggg tccctggacc tctaatcctc ttattttcga caactcatac
     5341 ttcacggagt tgttgagtgg tgagaaggaa ggtctccttc agctaccttc tgacaaggct
     5401 cttttgtctg accctgtatt ccgccctctc gttgacaaat atgcagcgga cgaagatgcc
     5461 ttctttgctg attacgctga ggctcaccaa aagctttccg agcttgggtt tgctgatgcc
     5521 gcggcgggag ggagcggacg ccagctttgg gactctgcgg tgagccactt cttttctctg
     5581 cccttcaaaa agagtctcct ggctgccttg atcctctgtc tcctggtggt gagatttgat
     5641 ccagcttccc cttcctccct gcacttcctc tacaagctgg cccagctctt ccgctggatc
     5701 cggaaggctg cattttctcg cctctaccag ctccgcatcc gtgactgagc ggccgctacg
     5761 taaattccgc ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg
     5821 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga
     5881 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa
     5941 tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa
     6001 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct
     6061 gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg
     6121 ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg
     6181 ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca
     6241 catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg
     6301 acgtggtttt cctttgaaaa acacgatgat aatatggcca caaccatggt taccgagtac
     6361 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     6421 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     6481 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     6541 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     6601 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     6661 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     6721 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     6781 ctccccggag tggaggcggc cgagcgcgcc ggggtgcc
//
