LOCUS       sequence-461304-        7235 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses Keima fused to PEX11B in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7235)
  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 7235)
  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-Pex11B
FEATURES             Location/Qualifiers
     source          1..7235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     236..256
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             510..979
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(1109..1126)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1280..1868)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1360..1379)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2032..2823,2824..2892))
                     /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     2655..2674
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2893..2997)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             3027..3620
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    3683..4040
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             4099..4515
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     4445..4467
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     4483..4499
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    4525..4899
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     4837..4853
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      4950..4959
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4956..4958,4959..4961,4962..5621)
                     /codon_start=1
                     /product="monomeric Keima-Red fluorescent protein, also 
                     known as mKeima"
                     /label=mKeima-Red
                     /translation="MVSVIAKQMTYKVYMSGTVNGHYFEVEGDGKGKPYEGEQTVKLTV
                     TKGGPLPFAWDILSPQLQYGSIPFTKYPEDIPDYFKQSFPEGYTWERSMNFEDGAVCTV
                     SNDSSIQGNCFIYNVKISGENFPPNGPVMQKKTQGWEPSTERLFARDGMLIGNDYMALK
                     LEGGGHYLCEFKSTYKAKKPVRMPGRHEIDRKLDVTSHNRDYTSVEQCEIAIARHSLLG
                     "
     misc_feature    6392..6958
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    6408..6960
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6552..6569)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6779..6798
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(6972..7235,1..339)
                     /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 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
       61 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
      121 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
      181 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
      241 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
      301 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag tcgacgataa aataaaagat
      361 tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct
      421 agcttaagta acgccatttt gcaaggcatg gaaaaataca taactgagaa tagagaagtt
      481 cagatcaagg tcaggaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt
      541 aagcagttcc tgccccggct cagggccaag aacagatgga acagctgaat atgggccaaa
      601 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca
      661 gatgcggtcc agccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa
      721 ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg
      781 ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca ctcggggcgc
      841 cagtcctccg attgactgag tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt
      901 tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc
      961 cgtcagcggg ggtctttcac atgcagcatg tatcaaaatt aatttggttt tttttcttaa
     1021 gtatttacat taaatggcca tagttgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     1081 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     1141 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     1201 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     1261 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     1321 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     1381 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     1441 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     1501 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     1561 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     1621 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     1681 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     1741 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     1801 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     1861 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     1921 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttgc
     1981 ggccggccgc aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     2041 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     2101 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     2161 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     2221 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     2281 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     2341 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     2401 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     2461 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     2521 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     2581 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     2641 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     2701 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     2761 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     2821 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     2881 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     2941 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     3001 atttccccga aaagtgccac ctgcataatg aaagacccca cctgtaggtt tggcaagcta
     3061 gcttaagtaa cgccattttg caaggcatgg aaaaatacat aactgagaat agaaaagttc
     3121 agatcaaggt caggaacaga tggaacagct gaatatgggc caaacaggat atctgtggta
     3181 agcagttcct gccccggctc agggccaaga acagatggaa cagctgaata tgggccaaac
     3241 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag
     3301 atgcggtcca gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag
     3361 gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt
     3421 tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc
     3481 agtcctccga ttgactgagt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt
     3541 gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc
     3601 gtcagcgggg gtctttcatt tgggggctcg tccgggatcg ggagacccct gcccagggac
     3661 caccgaccca ccaccgggag gtaagctggc cagcaactta tctgtgtctg tccgattgtc
     3721 tagtgtctat gactgatttt atgcgcctgc gtcggtacta gttagctaac tagctctgta
     3781 tctggcggac ccgtggtgga actgacgagt tcggaacacc cggccgcaac cctgggagac
     3841 gtcccaggga cttcgggggc cgtttttgtg gcccgacctg agtccaaaaa tcccgatcgt
     3901 tttggactct ttggtgcacc cccctaatag gagggatatg tggttctggt aggagacgag
     3961 aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggga ccgaagccgc
     4021 gccgcgcgtc ttgtctgctg cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg
     4081 tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac cttaggtcac
     4141 tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa gagacgttgg
     4201 gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg agacggcacc
     4261 tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg cccgcatgga
     4321 cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga cccccctccc
     4381 tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc cgccccgtct
     4441 ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc agccctcact
     4501 ccttctctag gcgcccccat atggccatat gagatcttat atggggcacc cccgcccctt
     4561 gtaaacttcc ctgaccctga catgacaaga gttactaaca gcccctctct ccaagctcac
     4621 ttacaggctc tctacttagt ccagcacgaa gtctggagac ctctggcggc agcctaccaa
     4681 gaacaactgg accgaccggt ggtacctcac ccttaccgag tcggcgacac agtgtgggtc
     4741 cgccgacacc agactaagaa cctagaacct cgctggaaag gaccttacac agtcctgctg
     4801 accaccccca ccgccctcaa agtagacggc atcgcagctt ggatacacgc cgcccacgtg
     4861 aaggctgccg accccggggg tggaccatcc tctagactgc cggatctagc tagttaatta
     4921 aggatcccag tgtggtggta cgggaattcg ccaccatggt gagtgtgatc gctaaacaaa
     4981 tgacctacaa ggtttatatg tcaggcacgg tcaatggaca ctactttgag gtcgaaggcg
     5041 atggaaaagg aaagccttac gagggagagc agacagtaaa gctcactgtc accaagggtg
     5101 gacctctgcc atttgcttgg gatattttat caccacagct tcagtacgga agcataccat
     5161 tcaccaagta ccctgaagac atccctgatt atttcaagca gtcattccct gagggatata
     5221 catgggagag gagcatgaac tttgaagatg gtgcagtgtg tactgtcagc aatgattcca
     5281 gcatccaagg caactgtttc atctacaatg tcaaaatctc tggtgagaac tttcctccca
     5341 atggacctgt tatgcagaag aagacacagg gctgggaacc cagcactgag cgtctctttg
     5401 cacgagatgg aatgctgata ggaaacgatt atatggctct gaagttggaa ggaggtggtc
     5461 actatttgtg tgaatttaaa tctacttaca aggcaaagaa gcctgtgagg atgccagggc
     5521 gccacgagat tgaccgcaaa ctggatgtaa ccagtcacaa cagggattac acatctgttg
     5581 agcagtgtga aatagccatt gcacgccact ctttgctcgg tggagggagc ggagacgcct
     5641 tcacccgctt caccaaccag acccagggcc gggaccgact cttcagagcc actcagtaca
     5701 catgcatgtt gcttagatat ttgttagagc ccaaagctgg caaagagaag gtggtaatga
     5761 agctcaagaa actggagtcc agtgtgagca ctggtcgtaa atggttcaga ctaggcaatg
     5821 tggtacatgc tatacaggca actgagcaga gcattcatgc cactgacctg gtacctcgct
     5881 tatgcttaac attagccaac ctgaaccgtg tgatttattt catctgtgac accatcctct
     5941 gggtgaggag cgtaggtctc acctctggca tcaacaaaga gaaatggcga acgagggctg
     6001 ctcaccacta ctactattct cttctgctga gcctggtcag ggatctgtat gaaatctccc
     6061 tgcagatgaa acgagttaca tgtgacaggg caaagaaaga gaaatcagca tcccaggatc
     6121 ctctttggtt cagcgtggct gaggaggaaa cagaatggct ccaatccttt ctacttcttt
     6181 tattccgatc tctgaagcag catcctccct tgctcctgga cacagtgaag aacctttgtg
     6241 atatcctgaa ccctttggac cagctgggga tctataaatc caatcctggc atcattggac
     6301 ttggaggtct tgtgtcctct atagcaggca tgatcactgt ggcatatcct cagatgaagc
     6361 tgaagacccg ttaggcggcc gctacgtaaa ttccgccccc ccccctaacg ttactggccg
     6421 aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc
     6481 gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag
     6541 gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt
     6601 tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa
     6661 ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc
     6721 aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg
     6781 gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat
     6841 gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa
     6901 acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataata
     6961 tggccacaac catggttacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
     7021 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
     7081 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
     7141 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
     7201 ggaccacgcc ggagagcgtc gaagcggggg cggtg
//
