LOCUS       sequence-403977-        7463 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses GPR35-mCherry in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7463)
  AUTHORS   Wyant GA, Yu W, Doulamis IP, Nomoto RS, Saeed MY, Duignan T, McCully
            JD, Kaelin WG Jr
  TITLE     Mitochondrial remodeling and ischemic protection by G 
            protein-coupled receptor 35 agonists.
  JOURNAL   Science. 2022 Aug 5;377(6606):621-629. doi: 10.1126/science.abm1638.
            Epub 2022 Aug 4.
  PUBMED    35926043
REFERENCE   2  (bases 1 to 7463)
  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"; doi: 
            "10.1126/science.abm1638"; journalName: "Science"; date: "2022-08-5-
            5"; volume: "377"; issue: "6606"; pages: "621-629"
COMMENT     Sequence Label: pMXSIP-GPR35-mCherry
FEATURES             Location/Qualifiers
     source          1..7463
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             22..615
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    678..1035
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1094..1510
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1440..1462
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1478..1494
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1520..1894
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1832..1848
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             2890..3597
                     /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(3032..3050)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3300..3319
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3615..4181
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3631..4183
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3775..3792)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4002..4021
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4195..4797
                     /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     4694..4714
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4968..5437
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5567..5584)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5738..6326)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5818..5837)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6490..7281,7282..7350))
                     /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"
     promoter        complement(7351..7455)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cccgaaaagt gccacctgca taatgaaaga ccccacctgt aggtttggca agctagctta
       61 agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaaa agttcagatc
      121 aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
      181 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat
      241 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
      301 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
      361 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
      421 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
      481 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
      541 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
      601 cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg
      661 acccaccacc gggaggtaag ctggccagca acttatctgt gtctgtccga ttgtctagtg
      721 tctatgactg attttatgcg cctgcgtcgg tactagttag ctaactagct ctgtatctgg
      781 cggacccgtg gtggaactga cgagttcgga acacccggcc gcaaccctgg gagacgtccc
      841 agggacttcg ggggccgttt ttgtggcccg acctgagtcc aaaaatcccg atcgttttgg
      901 actctttggt gcacccccct aataggaggg atatgtggtt ctggtaggag acgagaacct
      961 aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tgggaccgaa gccgcgccgc
     1021 gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt
     1081 gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa
     1141 agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac
     1201 cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa
     1261 ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc
     1321 agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt
     1381 caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc
     1441 ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc
     1501 tctaggcgcc cccatatggc catatgagat cttatatggg gcacccccgc cccttgtaaa
     1561 cttccctgac cctgacatga caagagttac taacagcccc tctctccaag ctcacttaca
     1621 ggctctctac ttagtccagc acgaagtctg gagacctctg gcggcagcct accaagaaca
     1681 actggaccga ccggtggtac ctcaccctta ccgagtcggc gacacagtgt gggtccgccg
     1741 acaccagact aagaacctag aacctcgctg gaaaggacct tacacagtcc tgctgaccac
     1801 ccccaccgcc ctcaaagtag acggcatcgc agcttggata cacgccgccc acgtgaaggc
     1861 tgccgacccc gggggtggac catcctctag actgccggat ctagctagtt aattaaggat
     1921 cccagtgtgg tggtacggga attcgccacc atgaatggca cctacaacac ctgtggctcc
     1981 agcgacctca cctggccccc agcgatcaag ctgggcttct acgcctactt gggcgtcctg
     2041 ctggtgctag gcctgctgct caacagcctg gcgctctggg tgttctgctg ccgcatgcag
     2101 cagtggacgg agacccgcat ctacatgacc aacctggcgg tggccgacct ctgcctgctg
     2161 tgcaccttgc ccttcgtgct gcactccctg cgagacacct cagacacgcc gctgtgccag
     2221 ctctcccagg gcatctacct gaccaacagg tacatgagca tcagcctggt cacggccatc
     2281 gccgtggacc gctatgtggc cgtgcggcac ccgctgcgtg cccgcgggct gcggtccccc
     2341 aggcaggctg cggccgtgtg cgcggtcctc tgggtgctgg tcatcggctc cctggtggct
     2401 cgctggctcc tggggattca ggagggcggc ttctgcttca ggagcacccg gcacaatttc
     2461 aactccatgg cgttcccgct gctgggattc tacctgcccc tggccgtggt ggtcttctgc
     2521 tccctgaagg tggtgactgc cctggcccag aggccaccca ccgacgtggg gcaggcagag
     2581 gccacccgca aggctgcccg catggtctgg gccaacctcc tggtgttcgt ggtctgcttc
     2641 ctgcccctgc acgtggggct gacagtgcgc ctcgcagtgg gctggaacgc ctgtgccctc
     2701 ctggagacga tccgtcgcgc cctgtacata accagcaagc tctcagatgc caactgctgc
     2761 ctggacgcca tctgctacta ctacatggcc aaggagttcc aggaggcgtc tgcactggcc
     2821 gtggctcccc gtgctaaggc ccacaaaagc caggactctc tgtgcgtgac cctcgccgga
     2881 gggagcggag tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
     2941 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     3001 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
     3061 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
     3121 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
     3181 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
     3241 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     3301 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
     3361 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
     3421 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     3481 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
     3541 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaggcg
     3601 gccgctacgt aaattccgcc ccccccccta acgttactgg ccgaagccgc ttggaataag
     3661 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     3721 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     3781 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     3841 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     3901 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     3961 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     4021 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     4081 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     4141 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatggtt
     4201 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc
     4261 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc
     4321 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     4381 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     4441 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     4501 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     4561 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     4621 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     4681 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     4741 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgagtc
     4801 gacgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac
     4861 ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata
     4921 actgagaata gagaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc
     4981 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac
     5041 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     5101 aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat
     5161 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     5221 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
     5281 acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc
     5341 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
     5401 tctgagtgat tgactacccg tcagcggggg tctttcacat gcagcatgta tcaaaattaa
     5461 tttggttttt tttcttaagt atttacatta aatggccata gttgcattaa tgaatcggcc
     5521 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5581 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5641 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5701 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5761 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5821 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5881 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5941 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6001 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6061 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6121 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6181 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6241 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6301 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6361 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6421 tcacctagat ccttttgcgg ccggccgcaa atcaatctaa agtatatatg agtaaacttg
     6481 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6541 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6601 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6661 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6721 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6781 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6841 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6901 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6961 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7021 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7081 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7141 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7201 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7261 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7321 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7381 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7441 aataggggtt ccgcgcacat ttc
//
