LOCUS       sequence-348092-        7056 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  knockout mouse Ldha.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7056)
  AUTHORS   Wu L, Hollinshead KER, Hao Y, Au C, Kroehling L, Ng C, Lin WY, Li D,
            Silva HM, Shin J, Lafaille JJ, Possemato R, Pacold ME, 
            Papagiannakopoulos T, Kimmelman AC, Satija R, Littman DR
  TITLE     Niche-Selective Inhibition of Pathogenic Th17 Cells by Targeting 
            Metabolic Redundancy.
  JOURNAL   Cell. 2020 Aug 6;182(3):641-654.e20. doi: 
            10.1016/j.cell.2020.06.014. Epub 2020 Jul 1.
  PUBMED    32615085
REFERENCE   2  (bases 1 to 7056)
  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: 
            "10.1016/j.cell.2020.06.014"; journalName: "Cell"; date: "2020-08-6-
            6"; volume: "182"; issue: "3"; pages: "641-654.e20"
COMMENT     Sequence Label: pSIN-Ldha
FEATURES             Location/Qualifiers
     source          1..7056
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     146..165
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(246..263)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(417..1005)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(497..516)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1176..1967,1968..2036))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1799..1818
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2037..2141)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2209..2227
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        2327..2706
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2707..2910
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2861..2881
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2911..3086
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    3149..3348
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             3549..3964
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3894..3916
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3932..3948
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        4011..4259
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4011..4031
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4182..4201
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        4280..4355
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4462..4673
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             4693..5178
                     /codon_start=1
                     /product="mouse GPI-anchored cell surface glycoprotein of 
                     the immunoglobulin superfamily"
                     /label=Thy1.1
                     /note="This allele of Thy1 is also known as CD90.1."
                     /translation="MNPAISVALLLSVLQVSRGQKVTSLTACLVNQNLRLDCRHENNTK
                     DNSIQHEFSLTREKRKHVLSGTLGIPEHTYRSRVTLSNQPYIKVLTLANFTTKDEGDYF
                     CELRVSGANPMSSNKSISVYRDKLVKCGGISLLVQNTSWMLLLLLSLSLLQALDFISL"
     CDS             5188..5244
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             5239..6039
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="GPMIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5299..5318)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5909..5928
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             6160..6585
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(6688..6710)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6846..6866)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6851..7056,1..124)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(7031..7056,1..110)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
       61 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
      121 caaaaagctt actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
      181 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc
      241 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
      301 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
      361 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      421 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      481 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      541 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      601 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      661 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      721 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      781 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
      841 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
      901 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
      961 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     1021 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     1081 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     1141 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     1201 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     1261 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     1321 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     1381 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     1441 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tgcaggcatc
     1501 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     1561 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     1621 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     1681 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     1741 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat
     1801 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     1861 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     1921 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     1981 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     2041 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     2101 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     2161 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     2221 acgaggccct ttcgtgttca agaattagct tggccattgc atacgttgta tccatatcat
     2281 aatatgtaca tttatattgg ctcatgtcca acattaccgc catgttgaca ttgattattg
     2341 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     2401 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     2461 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     2521 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     2581 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     2641 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     2701 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
     2761 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     2821 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
     2881 gtacggtggg aggtctatat aagcagagct caataaaaga gcccacaacc cctcactcgg
     2941 cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tattcccaat aaagcctctt
     3001 gctgtttgca tccgaatcgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg
     3061 actacccacg acgggggtct ttcatttggg ggctcgtccg ggatttggag acccctgccc
     3121 agggaccacc gacccaccac cgggaggtaa gctggccagc aacttatctg tgtctgtccg
     3181 attgtctagt gtctatgttt gatgttatgc gcctgcgtct gtactagtta gctaactagc
     3241 tctgtatctg gcggacccgt ggtggaactg acgagttctg aacacccggc cgcaaccctg
     3301 ggagacgtcc cagggacttt gggggccgtt tttgtggccc gacctgagga agggagtcga
     3361 tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg agacgagaac ctaaaacagt
     3421 tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg aagccgcgcg tcttgtctgc
     3481 tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg
     3541 aaaattaggg ccagactgtt accactccct taagtttgac cttaggtcac tggaaagatg
     3601 tcgagcggat cgctcacaac cagtcggtag atgtcaagaa gagacgttgg ttaccttctg
     3661 ctctgcagaa tggccaacct ttaacgtcgg atggccgcga gacggcacct ttaaccgaga
     3721 cctcatcacc caggttaaga tcaaggtctt ttcacctggc ccgcatggac acccagacca
     3781 ggtcccctac atcgtgacct gggaagcctt ggcttttgac ccccctccct gggtcaagcc
     3841 ctttgtacac cctaagcctc cgcctcctct tcctccatcc gccccgtctc tcccccttga
     3901 acctcctcgt tcgaccccgc ctcgatcctc cctttatcca gccctcactc cttctctagg
     3961 cgccggaatt gaagatcttt aattaacgat ctatcgattc tagatctaga gagggcctat
     4021 ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag ataattagaa
     4081 ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat
     4141 ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg
     4201 taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg
     4261 ctggtcatta tcaccgcggg ttttagagct agaaatagca agttaaaata aggctagtcc
     4321 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tgaatttaga tcttgagaca
     4381 aggtgccggg ccgggaattc ggcgaattgg gcccgacgtc gcatgctccc ggcccctcga
     4441 gtggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     4501 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
     4561 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     4621 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtgtcgt
     4681 gacgcggcca ccatgaaccc agccatcagc gtcgctctcc tgctctcagt cttgcaggtg
     4741 tcccgagggc agaaggtgac cagcctgaca gcctgcctgg tgaaccaaaa ccttcgcctg
     4801 gactgccgcc atgagaataa caccaaggat aactccatcc agcatgagtt cagcctgacc
     4861 cgagagaaga ggaagcacgt gctctcaggc accctcggga tacccgagca cacgtaccgc
     4921 tcccgcgtca ccctctccaa ccagccctat atcaaggtcc ttaccctagc caacttcacc
     4981 accaaggatg agggcgacta cttttgtgag cttcgagtct cgggcgcgaa tcccatgagc
     5041 tccaataaaa gtatcagtgt gtatagagac aaactggtca agtgtggcgg cataagcctg
     5101 ctggttcaga acacatcctg gatgctgctg ctgctgcttt ccctctccct cctccaagcc
     5161 ctggacttca tttctctggg cagcggcgcc accaacttca gcctgctgaa gcaggccggc
     5221 gacgtggagg agaaccccgg ccccatgatt gaacaagatg gattgcacgc aggttctccg
     5281 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     5341 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     5401 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     5461 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     5521 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     5581 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5641 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5701 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5761 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5821 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5881 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5941 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     6001 cgcatcgcct tctatcgcct tcttgacgag ttcttctgat tagatatcat cagtggtcca
     6061 ggctctagtt ttgactcaac aatatcacca gctgaagcct atagagtacg agccatagat
     6121 aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag
     6181 gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata cataactgag
     6241 aatagagaag ttcagatcaa ggtcaggaac agatggaaca gggtcgaccc tagagaacca
     6301 tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac
     6361 caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag
     6421 cccacaaccc ctcactcggg gcgccagtcc tccgattgac tgagtcgccc gggtacccgt
     6481 gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg
     6541 gtctcctctg agtgattgac tacccgtcag cgggggtctt tcatttgggg gctcgtccgg
     6601 gatcgggaga cccctgccca gggaccaccg acccaccacc gggaggtaag ctggctgcct
     6661 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     6721 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     6781 tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtagatccg
     6841 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     6901 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     6961 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     7021 aactccgccc atcccgcccc taactccgcc cagttc
//
