LOCUS       sequence-461303-        6704 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses Keima fused PEX26 peroxisomal localization sequence in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6704)
  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 6704)
  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 Keima-PEX26
FEATURES             Location/Qualifiers
     source          1..6704
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             244..713
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(843..860)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1014..1602)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1094..1113)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1766..2557,2558..2626))
                     /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     2389..2408
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2627..2731)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             2761..3354
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    3417..3774
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             3833..4249
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     4179..4201
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     4217..4233
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    4259..4633
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     4571..4587
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      4684..4693
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4690..4692,4693..4695,4696..5355)
                     /codon_start=1
                     /product="monomeric Keima-Red fluorescent protein, also 
                     known as mKeima"
                     /label=mKeima-Red
                     /translation="MVSVIAKQMTYKVYMSGTVNGHYFEVEGDGKGKPYEGEQTVKLTV
                     TKGGPLPFAWDILSPQLQYGSIPFTKYPEDIPDYFKQSFPEGYTWERSMNFEDGAVCTV
                     SNDSSIQGNCFIYNVKISGENFPPNGPVMQKKTQGWEPSTERLFARDGMLIGNDYMALK
                     LEGGGHYLCEFKSTYKAKKPVRMPGRHEIDRKLDVTSHNRDYTSVEQCEIAIARHSLLG
                     "
     misc_feature    5595..6161
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5611..6163
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5755..5772)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5982..6001
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(6175..6704,1..73)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MVTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     6674..6694
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
       61 gcccggtgcc tgagtcgacg ataaaataaa agattttatt tagtctccag aaaaaggggg
      121 gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg
      181 catggaaaaa tacataactg agaatagaga agttcagatc aaggtcagga acagatggaa
      241 cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc
      301 caagaacaga tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct
      361 gccccggctc agggccaaga acagatggtc cccagatgcg gtccagccct cagcagtttc
      421 tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat
      481 ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc
      541 aataaaagag cccacaaccc ctcactcggg gcgccagtcc tccgattgac tgagtcgccc
      601 gggtacccgt gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt
      661 ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt tcacatgcag
      721 catgtatcaa aattaatttg gttttttttc ttaagtattt acattaaatg gccatagttg
      781 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
      841 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      901 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      961 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1021 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     1081 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     1141 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     1201 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     1261 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     1321 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     1381 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     1441 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     1501 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     1561 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     1621 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     1681 ttatcaaaaa ggatcttcac ctagatcctt ttgcggccgg ccgcaaatca atctaaagta
     1741 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     1801 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     1861 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     1921 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     1981 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     2041 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     2101 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     2161 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     2221 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     2281 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     2341 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     2401 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     2461 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     2521 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     2581 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     2641 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     2701 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgcat
     2761 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
     2821 atggaaaaat acataactga gaatagaaaa gttcagatca aggtcaggaa cagatggaac
     2881 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     2941 aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg
     3001 ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct
     3061 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
     3121 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
     3181 ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgattgact gagtcgcccg
     3241 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc
     3301 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt catttggggg
     3361 ctcgtccggg atcgggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc
     3421 tggccagcaa cttatctgtg tctgtccgat tgtctagtgt ctatgactga ttttatgcgc
     3481 ctgcgtcggt actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac
     3541 gagttcggaa cacccggccg caaccctggg agacgtccca gggacttcgg gggccgtttt
     3601 tgtggcccga cctgagtcca aaaatcccga tcgttttgga ctctttggtg caccccccta
     3661 ataggaggga tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc
     3721 tgaatttttg ctttcggttt gggaccgaag ccgcgccgcg cgtcttgtct gctgcagcat
     3781 cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt agggccagac
     3841 tgttaccact cccttaagtt tgaccttagg tcactggaaa gatgtcgagc ggatcgctca
     3901 caaccagtcg gtagatgtca agaagagacg ttgggttacc ttctgctctg cagaatggcc
     3961 aacctttaac gtcggatggc cgcgagacgg cacctttaac cgagacctca tcacccaggt
     4021 taagatcaag gtcttttcac ctggcccgca tggacaccca gaccaggtcc cctacatcgt
     4081 gacctgggaa gccttggctt ttgacccccc tccctgggtc aagccctttg tacaccctaa
     4141 gcctccgcct cctcttcctc catccgcccc gtctctcccc cttgaacctc ctcgttcgac
     4201 cccgcctcga tcctcccttt atccagccct cactccttct ctaggcgccc ccatatggcc
     4261 atatgagatc ttatatgggg cacccccgcc ccttgtaaac ttccctgacc ctgacatgac
     4321 aagagttact aacagcccct ctctccaagc tcacttacag gctctctact tagtccagca
     4381 cgaagtctgg agacctctgg cggcagccta ccaagaacaa ctggaccgac cggtggtacc
     4441 tcacccttac cgagtcggcg acacagtgtg ggtccgccga caccagacta agaacctaga
     4501 acctcgctgg aaaggacctt acacagtcct gctgaccacc cccaccgccc tcaaagtaga
     4561 cggcatcgca gcttggatac acgccgccca cgtgaaggct gccgaccccg ggggtggacc
     4621 atcctctaga ctgccggatc tagctagtta attaaggatc ccagtgtggt ggtacgggaa
     4681 ttcgccacca tggtgagtgt gatcgctaaa caaatgacct acaaggttta tatgtcaggc
     4741 acggtcaatg gacactactt tgaggtcgaa ggcgatggaa aaggaaagcc ttacgaggga
     4801 gagcagacag taaagctcac tgtcaccaag ggtggacctc tgccatttgc ttgggatatt
     4861 ttatcaccac agcttcagta cggaagcata ccattcacca agtaccctga agacatccct
     4921 gattatttca agcagtcatt ccctgaggga tatacatggg agaggagcat gaactttgaa
     4981 gatggtgcag tgtgtactgt cagcaatgat tccagcatcc aaggcaactg tttcatctac
     5041 aatgtcaaaa tctctggtga gaactttcct cccaatggac ctgttatgca gaagaagaca
     5101 cagggctggg aacccagcac tgagcgtctc tttgcacgag atggaatgct gataggaaac
     5161 gattatatgg ctctgaagtt ggaaggaggt ggtcactatt tgtgtgaatt taaatctact
     5221 tacaaggcaa agaagcctgt gaggatgcca gggcgccacg agattgaccg caaactggat
     5281 gtaaccagtc acaacaggga ttacacatct gttgagcagt gtgaaatagc cattgcacgc
     5341 cactctttgc tcggtggagg gagcggacgc cagctttggg actctgcggt gagccacttc
     5401 ttttctctgc ccttcaaaaa gagtctcctg gctgccttga tcctctgtct cctggtggtg
     5461 agatttgatc cagcttcccc ttcctccctg cacttcctct acaagctggc ccagctcttc
     5521 cgctggatcc ggaaggctgc attttctcgc ctctaccagc tccgcatccg tgactgagcg
     5581 gccgctacgt aaattccgcc ccccccccta acgttactgg ccgaagccgc ttggaataag
     5641 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     5701 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     5761 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     5821 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     5881 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     5941 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     6001 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     6061 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     6121 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatggtt
     6181 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc
     6241 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc
     6301 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     6361 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     6421 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     6481 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     6541 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     6601 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     6661 acctccgcgc cccgcaacct ccccttctac gagcggctcg gctt
//
