LOCUS       sequence-461300-        6530 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses Keima fused to OMP25 mitochondrial localization sequence 
            in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6530)
  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 6530)
  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-OMP25
FEATURES             Location/Qualifiers
     source          1..6530
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(756..773)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(927..1515)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1007..1026)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1679..2470,2471..2539))
                     /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     2302..2321
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2540..2644)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             2674..3267
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    3330..3687
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             3746..4162
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     4092..4114
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     4130..4146
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    4172..4546
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     4484..4500
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      4597..4606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4603..4605,4606..4608,4609..5268)
                     /codon_start=1
                     /product="monomeric Keima-Red fluorescent protein, also 
                     known as mKeima"
                     /label=mKeima-Red
                     /translation="MVSVIAKQMTYKVYMSGTVNGHYFEVEGDGKGKPYEGEQTVKLTV
                     TKGGPLPFAWDILSPQLQYGSIPFTKYPEDIPDYFKQSFPEGYTWERSMNFEDGAVCTV
                     SNDSSIQGNCFIYNVKISGENFPPNGPVMQKKTQGWEPSTERLFARDGMLIGNDYMALK
                     LEGGGHYLCEFKSTYKAKKPVRMPGRHEIDRKLDVTSHNRDYTSVEQCEIAIARHSLLG
                     "
     misc_feature    5409..5975
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5425..5977
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5569..5586)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5796..5815
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(5989..6530,1..61)
                     /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     6488..6508
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
       61 agtcgacgat aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc
      121 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata
      181 cataactgag aatagagaag ttcagatcaa ggtcaggaac agatggaaca gctgaatatg
      241 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
      301 gtccccagat gcggtccagc cctcagcagt ttctagagaa ccatcagatg tttccagggt
      361 gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc
      421 gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc
      481 ggggcgccag tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc
      541 ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt
      601 gactacccgt cagcgggggt ctttcacatg cagcatgtat caaaattaat ttggtttttt
      661 ttcttaagta tttacattaa atggccatag ttgcattaat gaatcggcca acgcgcgggg
      721 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
      781 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
      841 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
      901 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
      961 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1021 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     1081 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     1141 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     1201 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     1261 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     1321 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     1381 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     1441 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     1501 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     1561 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     1621 cttttgcggc cggccgcaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     1681 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     1741 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     1801 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     1861 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     1921 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     1981 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     2041 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     2101 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     2161 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     2221 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     2281 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     2341 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     2401 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     2461 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     2521 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     2581 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     2641 cgcgcacatt tccccgaaaa gtgccacctg cataatgaaa gaccccacct gtaggtttgg
     2701 caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac tgagaataga
     2761 aaagttcaga tcaaggtcag gaacagatgg aacagctgaa tatgggccaa acaggatatc
     2821 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag ctgaatatgg
     2881 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
     2941 tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt ttccagggtg
     3001 ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg
     3061 cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg
     3121 gcgcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa taaaccctct
     3181 tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg
     3241 actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga gacccctgcc
     3301 cagggaccac cgacccacca ccgggaggta agctggccag caacttatct gtgtctgtcc
     3361 gattgtctag tgtctatgac tgattttatg cgcctgcgtc ggtactagtt agctaactag
     3421 ctctgtatct ggcggacccg tggtggaact gacgagttcg gaacacccgg ccgcaaccct
     3481 gggagacgtc ccagggactt cgggggccgt ttttgtggcc cgacctgagt ccaaaaatcc
     3541 cgatcgtttt ggactctttg gtgcaccccc ctaataggag ggatatgtgg ttctggtagg
     3601 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttgggaccg
     3661 aagccgcgcc gcgcgtcttg tctgctgcag catcgttctg tgttgtctct gtctgactgt
     3721 gtttctgtat ttgtctgaaa attagggcca gactgttacc actcccttaa gtttgacctt
     3781 aggtcactgg aaagatgtcg agcggatcgc tcacaaccag tcggtagatg tcaagaagag
     3841 acgttgggtt accttctgct ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga
     3901 cggcaccttt aaccgagacc tcatcaccca ggttaagatc aaggtctttt cacctggccc
     3961 gcatggacac ccagaccagg tcccctacat cgtgacctgg gaagccttgg cttttgaccc
     4021 ccctccctgg gtcaagccct ttgtacaccc taagcctccg cctcctcttc ctccatccgc
     4081 cccgtctctc ccccttgaac ctcctcgttc gaccccgcct cgatcctccc tttatccagc
     4141 cctcactcct tctctaggcg cccccatatg gccatatgag atcttatatg gggcaccccc
     4201 gccccttgta aacttccctg accctgacat gacaagagtt actaacagcc cctctctcca
     4261 agctcactta caggctctct acttagtcca gcacgaagtc tggagacctc tggcggcagc
     4321 ctaccaagaa caactggacc gaccggtggt acctcaccct taccgagtcg gcgacacagt
     4381 gtgggtccgc cgacaccaga ctaagaacct agaacctcgc tggaaaggac cttacacagt
     4441 cctgctgacc acccccaccg ccctcaaagt agacggcatc gcagcttgga tacacgccgc
     4501 ccacgtgaag gctgccgacc ccgggggtgg accatcctct agactgccgg atctagctag
     4561 ttaattaagg atcccagtgt ggtggtacgg gaattcgcca ccatggtgag tgtgatcgct
     4621 aaacaaatga cctacaaggt ttatatgtca ggcacggtca atggacacta ctttgaggtc
     4681 gaaggcgatg gaaaaggaaa gccttacgag ggagagcaga cagtaaagct cactgtcacc
     4741 aagggtggac ctctgccatt tgcttgggat attttatcac cacagcttca gtacggaagc
     4801 ataccattca ccaagtaccc tgaagacatc cctgattatt tcaagcagtc attccctgag
     4861 ggatatacat gggagaggag catgaacttt gaagatggtg cagtgtgtac tgtcagcaat
     4921 gattccagca tccaaggcaa ctgtttcatc tacaatgtca aaatctctgg tgagaacttt
     4981 cctcccaatg gacctgttat gcagaagaag acacagggct gggaacccag cactgagcgt
     5041 ctctttgcac gagatggaat gctgatagga aacgattata tggctctgaa gttggaagga
     5101 ggtggtcact atttgtgtga atttaaatct acttacaagg caaagaagcc tgtgaggatg
     5161 ccagggcgcc acgagattga ccgcaaactg gatgtaacca gtcacaacag ggattacaca
     5221 tctgttgagc agtgtgaaat agccattgca cgccactctt tgctcggtgg agggagcgga
     5281 cgaggcgacg gagagcctag tggagttcct gtagctgtgg tgctgctgcc agtgtttgcc
     5341 cttaccctgg tagcagtttg ggccttcgtg agataccgaa agcagctctg agcggccgct
     5401 acgtaaattc cgcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt
     5461 gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc
     5521 ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag
     5581 gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac
     5641 aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc
     5701 tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc
     5761 acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca
     5821 aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt
     5881 gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg
     5941 gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat ggttaccgag
     6001 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
     6061 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc
     6121 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     6181 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     6241 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     6301 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     6361 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     6421 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     6481 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
//
