LOCUS       sequence-349870-        6251 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of COX4 cDNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6251)
  AUTHORS   Spinelli JB, Rosen PC, Sprenger HG, Puszynska AM, Mann JL, Roessler 
            JM, Cangelosi AL, Henne A, Condon KJ, Zhang T, Kunchok T, Lewis CA, 
            Chandel NS, Sabatini DM
  TITLE     Fumarate is a terminal electron acceptor in the mammalian electron 
            transport chain.
  JOURNAL   Science. 2021 Dec 3;374(6572):1227-1237. doi: 
            10.1126/science.abi7495. Epub 2021 Dec 2.
  PUBMED    34855504
REFERENCE   2  (bases 1 to 6251)
  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.abi7495"; journalName: "Science"; date: "2021-12-3-
            3"; volume: "374"; issue: "6572"; pages: "1227-1237"
COMMENT     Sequence Label: pmxs_COX4
FEATURES             Location/Qualifiers
     source          1..6251
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(272..289)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(443..1031)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(523..542)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1195..1986,1987..2055))
                     /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     1818..1837
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2056..2160)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             2190..2783
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    2846..3203
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             3262..3678
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3608..3630
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3646..3662
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    3688..4062
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     4000..4016
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      4107..4116
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4682..4747
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    join(4771..5345,5346..5348,5349..5357)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4938..4955)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5165..5184
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5358..5753
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             join(5924..6251,1..142)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc
       61 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
      121 acccgtcagc gggggtcttt cacatgcagc atgtatcaaa attaatttgg ttttttttct
      181 taagtattta cattaaatgg ccatagttgc attaatgaat cggccaacgc gcggggagag
      241 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
      301 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
      361 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
      421 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
      481 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
      541 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
      601 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
      661 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
      721 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
      781 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
      841 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
      901 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
      961 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     1021 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     1081 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     1141 tgcggccggc cgcaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     1201 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1261 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     1321 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     1381 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1441 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     1501 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     1561 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     1621 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     1681 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1741 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     1801 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     1861 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1921 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     1981 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     2041 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     2101 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     2161 cacatttccc cgaaaagtgc cacctgcata atgaaagacc ccacctgtag gtttggcaag
     2221 ctagcttaag taacgccatt ttgcaaggca tggaaaaata cataactgag aatagaaaag
     2281 ttcagatcaa ggtcaggaac agatggaaca gctgaatatg ggccaaacag gatatctgtg
     2341 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga atatgggcca
     2401 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc
     2461 cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc
     2521 aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc
     2581 tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc
     2641 gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa ccctcttgca
     2701 gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta
     2761 cccgtcagcg ggggtctttc atttgggggc tcgtccggga tcgggagacc cctgcccagg
     2821 gaccaccgac ccaccaccgg gaggtaagct ggccagcaac ttatctgtgt ctgtccgatt
     2881 gtctagtgtc tatgactgat tttatgcgcc tgcgtcggta ctagttagct aactagctct
     2941 gtatctggcg gacccgtggt ggaactgacg agttcggaac acccggccgc aaccctggga
     3001 gacgtcccag ggacttcggg ggccgttttt gtggcccgac ctgagtccaa aaatcccgat
     3061 cgttttggac tctttggtgc acccccctaa taggagggat atgtggttct ggtaggagac
     3121 gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg ggaccgaagc
     3181 cgcgccgcgc gtcttgtctg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt
     3241 ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt
     3301 cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt
     3361 tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc
     3421 acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat
     3481 ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct
     3541 ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg
     3601 tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc
     3661 actccttctc taggcgcccc catatggcca tatgagatct tatatggggc acccccgccc
     3721 cttgtaaact tccctgaccc tgacatgaca agagttacta acagcccctc tctccaagct
     3781 cacttacagg ctctctactt agtccagcac gaagtctgga gacctctggc ggcagcctac
     3841 caagaacaac tggaccgacc ggtggtacct cacccttacc gagtcggcga cacagtgtgg
     3901 gtccgccgac accagactaa gaacctagaa cctcgctgga aaggacctta cacagtcctg
     3961 ctgaccaccc ccaccgccct caaagtagac ggcatcgcag cttggataca cgccgcccac
     4021 gtgaaggctg ccgaccccgg gggtggacca tcctctagac tgccggatct agctagttaa
     4081 ttaaggatcc gccacgccgt gttggtgcca ccatgggtat gggcctcgga gccagaatgt
     4141 tggctaccag ggtatttagc ctagttggca agcgagcaat ttccacctct gtgtgtgtac
     4201 gagctcatga aagtgttgtg aagagcgaag acttttcgct cccagcttat atggatcggc
     4261 gtgaccaccc cttgccggag gtggcccatg tcaagcacct gtctgccagc cagaaggcat
     4321 tgaaggagaa ggagaaggcc tcctggagca gcctctccat ggatgagaaa gtcgagttgt
     4381 atcgcattaa attcaaggag agctttgctg agatgaacag gggctcgaac gagtggaaga
     4441 cggttgtggg cggtgccatg ttcttcatcg gtttcaccgc gctcgttatc atgtggcaga
     4501 agcactatgt gtacggcccc ctcccgcaaa gctttgacaa agagtgggtg gccaagcaga
     4561 ccaagaggat gctggacatg aaggtgaacc ccatccaggg cttagcctcc aagtgggact
     4621 acgaaaagaa cgagtggaag aagtgagagc ggccgcagga ggcggtggag gcggtgcagg
     4681 cgactacaaa gaccatgacg gtgattataa agatcatgac attgattaca aggatgacga
     4741 tgacaagtaa taataaattc cgcccccccc cccctctccc tccccccccc ctaacgttac
     4801 tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat
     4861 attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat
     4921 tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga
     4981 agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca
     5041 gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac
     5101 acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt
     5161 caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca
     5221 ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt
     5281 aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg
     5341 ataatatggc cacaaccatg gttcctttgt ctcaagaaga atccaccctc attgaaagag
     5401 caacggctac aatcaacagc atccccatct ctgaagacta cagcgtcgcc agcgcagctc
     5461 tctctagcga cggccgcatc ttcactggtg tcaatgtata tcattttact gggggacctt
     5521 gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc agctggcaac ctgacttgta
     5581 tcgtcgcgat cggaaatgag aacaggggca tcttgagccc ctgcggacgg tgccgacagg
     5641 tgcttctcga tctgcatcct gggatcaaag ccatagtgaa ggacagtgat ggacagccga
     5701 cggcagttgg gattcgtgaa ttgctgccct ctggttatgt gtgggagggc taagtcgacg
     5761 ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt
     5821 aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg
     5881 agaatagaga agttcagatc aaggtcagga acagatggaa cagctgaata tgggccaaac
     5941 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggaacagct
     6001 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
     6061 acagatggtc cccagatgcg gtccagccct cagcagtttc tagagaacca tcagatgttt
     6121 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
     6181 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
     6241 ctcactcggg g
//
