LOCUS       sequence-461281-        7172 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses DEPP1 fused to mCherry in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7172)
  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 7172)
  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 DEPP1-mCherry
FEATURES             Location/Qualifiers
     source          1..7172
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    22..379
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             442..1035
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        1065..1169
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1170..1238,1239..2030)
                     /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     complement(1388..1407)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2194..2782
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2683..2702
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2936..2953
                     /label=L4440
                     /note="L4440 vector, forward primer"
     LTR             3083..3552
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(3723..4325)
                     /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     complement(3806..3826)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4337..4889
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4339..4905
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4499..4518)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     4728..4745
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(4923..5630)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(5201..5220)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5470..5488
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     misc_feature    6335..6709
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     complement(6381..6397)
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             complement(6719..7135)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(6735..6751)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(6767..6789)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
ORIGIN
        1 agacaacaca gaacgatgct gcagcagaca agacgcgcgg cgcggcttcg gtcccaaacc
       61 gaaagcaaaa attcagacgg aggcgggaac tgttttaggt tctcgtctcc taccagaacc
      121 acatatccct cctattaggg gggtgcacca aagagtccaa aacgatcggg atttttggac
      181 tcaggtcggg ccacaaaaac ggcccccgaa gtccctggga cgtctcccag ggttgcggcc
      241 gggtgttccg aactcgtcag ttccaccacg ggtccgccag atacagagct agttagctaa
      301 ctagtaccga cgcaggcgca taaaatcagt catagacact agacaatcgg acagacacag
      361 ataagttgct ggccagctta cctcccggtg gtgggtcggt ggtccctggg caggggtctc
      421 ccgatcccgg acgagccccc aaatgaaaga cccccgctga cgggtagtca atcactcaga
      481 ggagaccctc ccaaggaaca gcgagaccac aagtcggatg caactgcaag agggtttatt
      541 ggatacacgg gtacccgggc gactcagtca atcggaggac tggcgcgccg agtgaggggt
      601 tgtgggctct tttattgagc tcggggagca gaagcgcgcg aacagaagcg agaagcgaac
      661 tgattggtta gttcaaataa ggcacagggt catttcaggt ccttggggca ccctggaaac
      721 atctgatggt tctctagaaa ctgctgaggg ctggaccgca tctggggacc atctgttctt
      781 ggccctgagc cggggcagga actgcttacc acagatatcc tgtttggccc atattcagct
      841 gttccatctg ttcttggccc tgagccgggg caggaactgc ttaccacaga tatcctgttt
      901 ggcccatatt cagctgttcc atctgttcct gaccttgatc tgaacttttc tattctcagt
      961 tatgtatttt tccatgcctt gcaaaatggc gttacttaag ctagcttgcc aaacctacag
     1021 gtggggtctt tcattatgca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     1081 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     1141 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     1201 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     1261 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     1321 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     1381 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     1441 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     1501 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     1561 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     1621 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     1681 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     1741 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     1801 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     1861 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     1921 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     1981 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     2041 aagtttactc atatatactt tagattgatt tgcggccggc cgcaaaagga tctaggtgaa
     2101 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     2161 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2221 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2281 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2341 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2401 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2461 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2521 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2581 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2641 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2701 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     2761 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     2821 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     2881 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     2941 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     3001 gccgattcat taatgcaact atggccattt aatgtaaata cttaagaaaa aaaaccaaat
     3061 taattttgat acatgctgca tgtgaaagac ccccgctgac gggtagtcaa tcactcagag
     3121 gagaccctcc caaggaacag cgagaccaca agtcggatgc aactgcaaga gggtttattg
     3181 gatacacggg tacccgggcg actcagtcaa tcggaggact ggcgcgccga gtgaggggtt
     3241 gtgggctctt ttattgagct cggggagcag aagcgcgcga acagaagcga gaagcgaact
     3301 gattggttag ttcaaataag gcacagggtc atttcaggtc cttggggcac cctggaaaca
     3361 tctgatggtt ctctagaaac tgctgagggc tggaccgcat ctggggacca tctgttcttg
     3421 gccctgagcc ggggcaggaa ctgcttacca cagatatcct gtttggccca tattcagctg
     3481 ttccatctgt tcttggccct gagccggggc aggaactgct taccacagat atcctgtttg
     3541 gcccatattc agctgttcca tctgttcctg accttgatct gaacttctct attctcagtt
     3601 atgtattttt ccatgccttg caaaatggcg ttacttaagc tagcttgcca aacctacagg
     3661 tggggtcttt cattcccccc tttttctgga gactaaataa aatcttttat tttatcgtcg
     3721 actcaggcac cgggcttgcg ggtcatgcac caggtgcgcg gtccttcggg cacctcgacg
     3781 tcggcggtga cggtgaagcc gagccgctcg tagaagggga ggttgcgggg cgcggaggtc
     3841 tccaggaagg cgggcacccc ggcgcgctcg gccgcctcca ctccggggag cacgacggcg
     3901 ctgcccagac ccttgccctg gtggtcgggc gagacgccga cggtggccag gaaccacgcg
     3961 ggctccttgg gccggtgcgg cgccaggagg ccttccatct gttgctgcgc ggccagccgg
     4021 gaaccgctca actcggccat gcgcgggccg atctcggcga acaccgcccc cgcttcgacg
     4081 ctctccggcg tggtccagac cgccaccgcg gcgccgtcgt ccgcgaccca caccttgccg
     4141 atgtcgagcc cgacgcgcgt gaggaagagt tcttgcagct cggtgacccg ctcgatgtgg
     4201 cggtccggat cgacggtgtg gcgcgtggcg gggtagtcgg cgaacgcggc ggcgagggtg
     4261 cgtacggccc tggggacgtc gtcgcgggtg gcgaggcgca ccgtgggctt gtactcggta
     4321 accatggttg tggccatatt atcatcgtgt ttttcaaagg aaaaccacgt ccccgtggtt
     4381 cggggggcct agacgttttt ttaacctcga ctaaacacat gtaaagcatg tgcaccgagg
     4441 ccccagatca gatcccatac aatggggtac cttctgggca tccttcagcc ccttgttgaa
     4501 tacgcttgag gagagccatt tgactctttc cacaactatc caactcacaa cgtggcactg
     4561 gggttgtgcc gcctttgcag gtgtatctta tacacgtggc ttttggccgc agaggcacct
     4621 gtcgccaggt ggggggttcc gctgcctgca aagggtcgct acagacgttg tttgtcttca
     4681 agaagcttcc agaggaactg cttccttcac gacattcaac agaccttgca ttcctttggc
     4741 gagaggggaa agacccctag gaatgctcgt caagaagaca gggccaggtt tccgggccct
     4801 cacattgcca aaagacggca atatggtgga aaataacata tagacaaacg cacaccggcc
     4861 ttattccaag cggcttcggc cagtaacgtt aggggggggg gcggaattta cgtagcggcc
     4921 gcctacttgt acagctcgtc catgccgccg gtggagtggc ggccctcggc gcgttcgtac
     4981 tgttccacga tggtgtagtc ctcgttgtgg gaggtgatgt ccaacttgat gttgacgttg
     5041 taggcgccgg gcagctgcac gggcttcttg gccttgtagg tggtcttgac ctcagcgtcg
     5101 tagtggccgc cgtccttcag cttcagcctc tgcttgatct cgcccttcag ggcgccgtcc
     5161 tcggggtaca tccgctcgga ggaggcctcc cagcccatgg tcttcttctg cattacgggg
     5221 ccgtcggagg ggaagttggt gccgcgcagc ttcaccttgt agatgaactc gccgtcctgc
     5281 agggaggagt cctgggtcac ggtcaccacg ccgccgtcct cgaagttcat cacgcgctcc
     5341 cacttgaagc cctcggggaa ggacagcttc aagtagtcgg ggatgtcggc ggggtgcttc
     5401 acgtaggcct tggagccgta catgaactga ggggacagga tgtcccaggc gaagggcagg
     5461 gggccaccct tggtcacctt cagcttggcg gtctgggtgc cctcgtaggg gcggccctcg
     5521 ccctcgccct cgatctcgaa ctcgtggccg ttcacggagc cctccatgtg caccttgaag
     5581 cgcatgaact ccttgatgat ggccatgtta tcctcctcgc ccttgctcac tccgctccct
     5641 ccgagttcat ggatcaccgg gaggtgcgag tagagtgttc tgaggacact gctggggcgg
     5701 gggcggtggc gggaggccat ggcttgggca gactgcccaa aagtccagct tcttaggtca
     5761 gagctctgct gcccatcctg cggtggtctt gccagggaat gcccaggcat cctggcttca
     5821 cagagcctcc ctctgggggc ccccatgggc ttgctgctgt ccatctgtct atgtggaccc
     5881 cagaggccag cagaggggcc agtcctcctt ggcaggtccc tctggcttgg ctgcttttcc
     5941 tgggactccc caaaaagcca gtccaggggg tccacagtat cagccatggg cagtgtgggc
     6001 tgctggccac tgaaacgtgc ggtgatgtct cgcagggaca cagcagggcg gcccttccgg
     6061 caggcctggg ctggcctgtg gggtggcctg ggttggccct gggccgtggc cttgtccagc
     6121 acagaggtgg gctgtgccag tcgagatata gacctcacgt agtcatccag gctaggagag
     6181 ggtgggggct cctgtccagg acccccaagc agcatctcct ccgtggtctc ccgaattgtg
     6241 ggcagatggg ccacggagag cagaagccgg gacctcatgg tggcgaattc ccgtaccacc
     6301 acactgggat ccttaattaa ctagctagat ccggcagtct agaggatggt ccacccccgg
     6361 ggtcggcagc cttcacgtgg gcggcgtgta tccaagctgc gatgccgtct actttgaggg
     6421 cggtgggggt ggtcagcagg actgtgtaag gtcctttcca gcgaggttct aggttcttag
     6481 tctggtgtcg gcggacccac actgtgtcgc cgactcggta agggtgaggt accaccggtc
     6541 ggtccagttg ttcttggtag gctgccgcca gaggtctcca gacttcgtgc tggactaagt
     6601 agagagcctg taagtgagct tggagagagg ggctgttagt aactcttgtc atgtcagggt
     6661 cagggaagtt tacaaggggc gggggtgccc catataagat ctcatatggc catatggggg
     6721 cgcctagaga aggagtgagg gctggataaa gggaggatcg aggcggggtc gaacgaggag
     6781 gttcaagggg gagagacggg gcggatggag gaagaggagg cggaggctta gggtgtacaa
     6841 agggcttgac ccagggaggg gggtcaaaag ccaaggcttc ccaggtcacg atgtagggga
     6901 cctggtctgg gtgtccatgc gggccaggtg aaaagacctt gatcttaacc tgggtgatga
     6961 ggtctcggtt aaaggtgccg tctcgcggcc atccgacgtt aaaggttggc cattctgcag
     7021 agcagaaggt aacccaacgt ctcttcttga catctaccga ctggttgtga gcgatccgct
     7081 cgacatcttt ccagtgacct aaggtcaaac ttaagggagt ggtaacagtc tggccctaat
     7141 tttcagacaa atacagaaac acagtcagac ag
//
