LOCUS       sequence-461301-        7174 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses DEPP1 fused to mNEONGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7174)
  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 7174)
  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-mNG
FEATURES             Location/Qualifiers
     source          1..7174
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     205..225
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             479..948
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(1078..1095)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1249..1837)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1329..1348)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2001..2792,2793..2861))
                     /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     2624..2643
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2862..2966)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             2996..3589
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    3652..4009
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             4068..4484
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     4414..4436
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     4452..4468
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    4494..4868
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     4806..4822
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             5573..6277
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="VSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYEE
                     LNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFEDG
                     ASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIST
                     FKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQK
                     AFTDVMGMDELYK"
     regulatory      5825..5834
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    6300..6866
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    6316..6868
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6460..6477)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6687..6706
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(6880..7174,1..308)
                     /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 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc
       61 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg
      121 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg
      181 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg
      241 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg
      301 gtgcctgagt cgacgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg
      361 aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg
      421 aaaaatacat aactgagaat agagaagttc agatcaaggt caggaacaga tggaacagct
      481 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      541 acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
      601 ggctcagggc caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagag
      661 aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa
      721 ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa
      781 aagagcccac aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta
      841 cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg
      901 ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcaca tgcagcatgt
      961 atcaaaatta atttggtttt ttttcttaag tatttacatt aaatggccat agttgcatta
     1021 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     1081 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     1141 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     1201 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1261 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1321 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1381 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1441 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1501 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1561 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1621 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1681 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     1741 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     1801 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     1861 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     1921 aaaaaggatc ttcacctaga tccttttgcg gccggccgca aatcaatcta aagtatatat
     1981 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2041 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     2101 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     2161 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     2221 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     2281 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     2341 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     2401 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     2461 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     2521 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     2581 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     2641 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     2701 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     2761 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     2821 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     2881 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     2941 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgcataatga
     3001 aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga
     3061 aaaatacata actgagaata gaaaagttca gatcaaggtc aggaacagat ggaacagctg
     3121 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     3181 cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     3241 gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga
     3301 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
     3361 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa
     3421 agagcccaca acccctcact cggcgcgcca gtcctccgat tgactgagtc gcccgggtac
     3481 ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg
     3541 gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt gggggctcgt
     3601 ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg taagctggcc
     3661 agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta tgcgcctgcg
     3721 tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt
     3781 cggaacaccc ggccgcaacc ctgggagacg tcccagggac ttcgggggcc gtttttgtgg
     3841 cccgacctga gtccaaaaat cccgatcgtt ttggactctt tggtgcaccc ccctaatagg
     3901 agggatatgt ggttctggta ggagacgaga acctaaaaca gttcccgcct ccgtctgaat
     3961 ttttgctttc ggtttgggac cgaagccgcg ccgcgcgtct tgtctgctgc agcatcgttc
     4021 tgtgttgtct ctgtctgact gtgtttctgt atttgtctga aaattagggc cagactgtta
     4081 ccactccctt aagtttgacc ttaggtcact ggaaagatgt cgagcggatc gctcacaacc
     4141 agtcggtaga tgtcaagaag agacgttggg ttaccttctg ctctgcagaa tggccaacct
     4201 ttaacgtcgg atggccgcga gacggcacct ttaaccgaga cctcatcacc caggttaaga
     4261 tcaaggtctt ttcacctggc ccgcatggac acccagacca ggtcccctac atcgtgacct
     4321 gggaagcctt ggcttttgac ccccctccct gggtcaagcc ctttgtacac cctaagcctc
     4381 cgcctcctct tcctccatcc gccccgtctc tcccccttga acctcctcgt tcgaccccgc
     4441 ctcgatcctc cctttatcca gccctcactc cttctctagg cgcccccata tggccatatg
     4501 agatcttata tggggcaccc ccgccccttg taaacttccc tgaccctgac atgacaagag
     4561 ttactaacag cccctctctc caagctcact tacaggctct ctacttagtc cagcacgaag
     4621 tctggagacc tctggcggca gcctaccaag aacaactgga ccgaccggtg gtacctcacc
     4681 cttaccgagt cggcgacaca gtgtgggtcc gccgacacca gactaagaac ctagaacctc
     4741 gctggaaagg accttacaca gtcctgctga ccacccccac cgccctcaaa gtagacggca
     4801 tcgcagcttg gatacacgcc gcccacgtga aggctgccga ccccgggggt ggaccatcct
     4861 ctagactgcc ggatctagct agttaattaa ggatcccagt gtggtggtac gggaattcgc
     4921 caccatgagg tcccggcttc tgctctccgt ggcccatctg cccacaattc gggagaccac
     4981 ggaggagatg ctgcttgggg gtcctggaca ggagccccca ccctctccta gcctggatga
     5041 ctacgtgagg tctatatctc gactggcaca gcccacctct gtgctggaca aggccacggc
     5101 ccagggccaa cccaggccac cccacaggcc agcccaggcc tgccggaagg gccgccctgc
     5161 tgtgtccctg cgagacatca ccgcacgttt cagtggccag cagcccacac tgcccatggc
     5221 tgatactgtg gaccccctgg actggctttt tggggagtcc caggaaaagc agccaagcca
     5281 gagggacctg ccaaggagga ctggcccctc tgctggcctc tggggtccac atagacagat
     5341 ggacagcagc aagcccatgg gggcccccag agggaggctc tgtgaagcaa ggatgcctgg
     5401 gcattccctg gcaagaccac cgcaggatgg gcagcagagc tctgacctaa gaagctggac
     5461 ttttgggcag tctgcccaag ccatggcctc ccgccaccgc ccccgcccca gcagtgtcct
     5521 cagaacactc tactcgcacc tcccggtgat ccatgaactc ggagggagcg gagtgagcaa
     5581 gggcgaggag gataacatgg cctctctccc agcgacacat gagttacaca tctttggctc
     5641 catcaacggt gtggactttg acatggtggg tcagggcacc ggcaatccaa atgatggtta
     5701 tgaggagtta aacctgaagt ccaccaaggg tgacctccag ttctccccct ggattctggt
     5761 ccctcatatc gggtatggct tccatcagta cctgccctac cctgacggga tgtcgccttt
     5821 ccaggccgcc atggtagatg gctccggata ccaagtccat cgcacaatgc agtttgaaga
     5881 tggtgcctcc cttactgtta actaccgcta cacctacgag ggaagccaca tcaaaggaga
     5941 ggcccaggtg aaggggactg gtttccctgc tgacggtcct gtgatgacca actcgctgac
     6001 cgctgcggac tggtgcaggt cgaagaagac ttaccccaac gacaaaacca tcatcagtac
     6061 ctttaagtgg agttacacca ctggaaatgg caagcgctac cggagcactg cgcggaccac
     6121 ctacaccttt gccaagccaa tggcggctaa ctatctgaag aaccagccga tgtacgtgtt
     6181 ccgtaagacg gagctcaagc actccaagac cgagctcaac ttcaaggagt ggcaaaaggc
     6241 ctttaccgat gtgatgggca tggacgagct gtacaagtat aggcggccgc tacgtaaatt
     6301 ccgccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
     6361 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     6421 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     6481 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt
     6541 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
     6601 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     6661 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
     6721 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct
     6781 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     6841 ttttcctttg aaaaacacga tgataatatg gccacaacca tggttaccga gtacaagccc
     6901 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     6961 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     7021 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     7081 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     7141 gtgttcgccg agatcggccc gcgcatggcc gagt
//
