LOCUS       sequence-461248-        6986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses DEPP1 dTsnare fused to mCherry in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6986)
  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 6986)
  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 dTSnare-mCherry
FEATURES             Location/Qualifiers
     source          1..6986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             349..942
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        972..1076
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1077..1145,1146..1937)
                     /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(1295..1314)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2101..2689
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2590..2609
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2843..2860
                     /label=L4440
                     /note="L4440 vector, forward primer"
     LTR             2990..3459
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(3630..4232)
                     /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(3713..3733)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4244..4796
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4246..4812
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4406..4425)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     4635..4652
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(4830..5537)
                     /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(5108..5127)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5377..5395
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     misc_feature    6056..6430
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     complement(6102..6118)
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             complement(6440..6856)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(6456..6472)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(6488..6510)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    join(6915..6986,1..286)
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 tttaggttct cgtctcctac cagaaccaca tatccctcct attagggggg tgcaccaaag
       61 agtccaaaac gatcgggatt tttggactca ggtcgggcca caaaaacggc ccccgaagtc
      121 cctgggacgt ctcccagggt tgcggccggg tgttccgaac tcgtcagttc caccacgggt
      181 ccgccagata cagagctagt tagctaacta gtaccgacgc aggcgcataa aatcagtcat
      241 agacactaga caatcggaca gacacagata agttgctggc cagcttacct cccggtggtg
      301 ggtcggtggt ccctgggcag gggtctcccg atcccggacg agcccccaaa tgaaagaccc
      361 ccgctgacgg gtagtcaatc actcagagga gaccctccca aggaacagcg agaccacaag
      421 tcggatgcaa ctgcaagagg gtttattgga tacacgggta cccgggcgac tcagtcaatc
      481 ggaggactgg cgcgccgagt gaggggttgt gggctctttt attgagctcg gggagcagaa
      541 gcgcgcgaac agaagcgaga agcgaactga ttggttagtt caaataaggc acagggtcat
      601 ttcaggtcct tggggcaccc tggaaacatc tgatggttct ctagaaactg ctgagggctg
      661 gaccgcatct ggggaccatc tgttcttggc cctgagccgg ggcaggaact gcttaccaca
      721 gatatcctgt ttggcccata ttcagctgtt ccatctgttc ttggccctga gccggggcag
      781 gaactgctta ccacagatat cctgtttggc ccatattcag ctgttccatc tgttcctgac
      841 cttgatctga acttttctat tctcagttat gtatttttcc atgccttgca aaatggcgtt
      901 acttaagcta gcttgccaaa cctacaggtg gggtctttca ttatgcaggt ggcacttttc
      961 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     1021 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     1081 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     1141 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     1201 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     1261 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     1321 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     1381 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     1441 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     1501 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     1561 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     1621 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     1681 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     1741 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     1801 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     1861 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     1921 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttgc
     1981 ggccggccgc aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     2041 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2101 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2161 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2221 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     2281 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2341 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2401 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2461 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2521 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2581 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2641 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2701 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     2761 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     2821 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     2881 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcaactatg gccatttaat
     2941 gtaaatactt aagaaaaaaa accaaattaa ttttgataca tgctgcatgt gaaagacccc
     3001 cgctgacggg tagtcaatca ctcagaggag accctcccaa ggaacagcga gaccacaagt
     3061 cggatgcaac tgcaagaggg tttattggat acacgggtac ccgggcgact cagtcaatcg
     3121 gaggactggc gccccgagtg aggggttgtg ggctctttta ttgagctcgg ggagcagaag
     3181 cgcgcgaaca gaagcgagaa gcgaactgat tggttagttc aaataaggca cagggtcatt
     3241 tcaggtcctt ggggcaccct ggaaacatct gatggttctc tagaaactgc tgagggctgg
     3301 accgcatctg gggaccatct gttcttggcc ctgagccggg gcaggaactg cttaccacag
     3361 atatcctgtt tggcccatat tcagctgttc catctgttct tggccctgag ccggggcagg
     3421 aactgcttac cacagatatc ctgtttggcc catattcagc tgttccatct gttcctgacc
     3481 ttgatctgaa cttctctatt ctcagttatg tatttttcca tgccttgcaa aatggcgtta
     3541 cttaagctag cttgccaaac ctacaggtgg ggtctttcat tccccccttt ttctggagac
     3601 taaataaaat cttttatttt atcgtcgact caggcaccgg gcttgcgggt catgcaccag
     3661 gtgcgcggtc cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag
     3721 aaggggaggt tgcggggcgc ggaggtctcc aggaaggcgg gcaccccggc gcgctcggcc
     3781 gcctccactc cggggagcac gacggcgctg cccagaccct tgccctggtg gtcgggcgag
     3841 acgccgacgg tggccaggaa ccacgcgggc tccttgggcc ggtgcggcgc caggaggcct
     3901 tccatctgtt gctgcgcggc cagccgggaa ccgctcaact cggccatgcg cgggccgatc
     3961 tcggcgaaca ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg
     4021 ccgtcgtccg cgacccacac cttgccgatg tcgagcccga cgcgcgtgag gaagagttct
     4081 tgcagctcgg tgacccgctc gatgtggcgg tccggatcga cggtgtggcg cgtggcgggg
     4141 tagtcggcga acgcggcggc gagggtgcgt acggccctgg ggacgtcgtc gcgggtggcg
     4201 aggcgcaccg tgggcttgta ctcggtaacc atggttgtgg ccatattatc atcgtgtttt
     4261 tcaaaggaaa accacgtccc cgtggttcgg ggggcctaga cgttttttta acctcgacta
     4321 aacacatgta aagcatgtgc accgaggccc cagatcagat cccatacaat ggggtacctt
     4381 ctgggcatcc ttcagcccct tgttgaatac gcttgaggag agccatttga ctctttccac
     4441 aactatccaa ctcacaacgt ggcactgggg ttgtgccgcc tttgcaggtg tatcttatac
     4501 acgtggcttt tggccgcaga ggcacctgtc gccaggtggg gggttccgct gcctgcaaag
     4561 ggtcgctaca gacgttgttt gtcttcaaga agcttccaga ggaactgctt ccttcacgac
     4621 attcaacaga ccttgcattc ctttggcgag aggggaaaga cccctaggaa tgctcgtcaa
     4681 gaagacaggg ccaggtttcc gggccctcac attgccaaaa gacggcaata tggtggaaaa
     4741 taacatatag acaaacgcac accggcctta ttccaagcgg cttcggccag taacgttagg
     4801 ggggggggcg gaatttacgt agcggccgcc tacttgtaca gctcgtccat gccgccggtg
     4861 gagtggcggc cctcggcgcg ttcgtactgt tccacgatgg tgtagtcctc gttgtgggag
     4921 gtgatgtcca acttgatgtt gacgttgtag gcgccgggca gctgcacggg cttcttggcc
     4981 ttgtaggtgg tcttgacctc agcgtcgtag tggccgccgt ccttcagctt cagcctctgc
     5041 ttgatctcgc ccttcagggc gccgtcctcg gggtacatcc gctcggagga ggcctcccag
     5101 cccatggtct tcttctgcat tacggggccg tcggagggga agttggtgcc gcgcagcttc
     5161 accttgtaga tgaactcgcc gtcctgcagg gaggagtcct gggtcacggt caccacgccg
     5221 ccgtcctcga agttcatcac gcgctcccac ttgaagccct cggggaagga cagcttcaag
     5281 tagtcgggga tgtcggcggg gtgcttcacg taggccttgg agccgtacat gaactgaggg
     5341 gacaggatgt cccaggcgaa gggcaggggg ccacccttgg tcaccttcag cttggcggtc
     5401 tgggtgccct cgtaggggcg gccctcgccc tcgccctcga tctcgaactc gtggccgttc
     5461 acggagccct ccatgtgcac cttgaagcgc atgaactcct tgatgatggc catgttatcc
     5521 tcctcgccct tgctcactcc gctccctccg agttcatgga tcaccgggag gtgcgagtag
     5581 agtgttctga ggacactgct ggggcggggg cggtggcggg aggccatggc ttgggcagac
     5641 tgcccaaaag tccagcttct taggtcagag ctctgctgcc catcctgcgg tggtcttgcc
     5701 agggaatgcc caggcatcct ggcttcacag agcctccctc tgggggcccc catgggcttg
     5761 ctgctgtcca tctgtctatg tggaccccag aggccagcag aggggccagt cctccttggc
     5821 aggtccctct ggcttggctg cttttcctgg gactccccaa aaagccagtc cagggggtcc
     5881 acagtatcag ccatgggcag tgtgggctgc tggccactga aacgtgcggt gatgtctcgc
     5941 agggacacag cagggcggcc cttccggcag gcctgggctg gcctgtgggg tggcctcatg
     6001 gtggcgaatt cccgtaccac cacactggga tccttaatta actagctaga tccggcagtc
     6061 tagaggatgg tccacccccg gggtcggcag ccttcacgtg ggcggcgtgt atccaagctg
     6121 cgatgccgtc tactttgagg gcggtggggg tggtcagcag gactgtgtaa ggtcctttcc
     6181 agcgaggttc taggttctta gtctggtgtc ggcggaccca cactgtgtcg ccgactcggt
     6241 aagggtgagg taccaccggt cggtccagtt gttcttggta ggctgccgcc agaggtctcc
     6301 agacttcgtg ctggactaag tagagagcct gtaagtgagc ttggagagag gggctgttag
     6361 taactcttgt catgtcaggg tcagggaagt ttacaagggg cgggggtgcc ccatataaga
     6421 tctcatatgg ccatatgggg gcgcctagag aaggagtgag ggctggataa agggaggatc
     6481 gaggcggggt cgaacgagga ggttcaaggg ggagagacgg ggcggatgga ggaagaggag
     6541 gcggaggctt agggtgtaca aagggcttga cccagggagg ggggtcaaaa gccaaggctt
     6601 cccaggtcac gatgtagggg acctggtctg ggtgtccatg cgggccaggt gaaaagacct
     6661 tgatcttaac ctgggtgatg aggtctcggt taaaggtgcc gtctcgcggc catccgacgt
     6721 taaaggttgg ccattctgca gagcagaagg taacccaacg tctcttcttg acatctaccg
     6781 actggttgtg agcgatccgc tcgacatctt tccagtgacc taaggtcaaa cttaagggag
     6841 tggtaacagt ctggccctaa ttttcagaca aatacagaaa cacagtcaga cagagacaac
     6901 acagaacgat gctgcagcag acaagacgcg cggcgcggct tcggtcccaa accgaaagca
     6961 aaaattcaga cggaggcggg aactgt
//
