LOCUS       sequence-348096-        7056 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  knockout mouse CD45.
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-CD45
FEATURES             Location/Qualifiers
     source          1..7056
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     156..175
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(256..273)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(427..1015)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(507..526)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1186..1977,1978..2046))
                     /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     1809..1828
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2047..2151)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2219..2237
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        2337..2716
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2717..2920
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2871..2891
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2921..3096
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    3159..3358
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             3559..3974
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3904..3926
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3942..3958
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        4021..4269
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4021..4041
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4192..4211
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        4290..4365
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4472..4683
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             4703..5188
                     /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             5198..5254
                     /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             5249..6049
                     /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(5309..5328)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5919..5938
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             6170..6595
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(6698..6720)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6856..6876)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6861..7056,1..134)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(7041..7056,1..120)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
       61 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
      121 taggcttttg caaaaagctt actggcttaa ctatgcggca tcagagcaga ttgtactgag
      181 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
      241 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
      301 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
      361 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
      421 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
      481 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
      541 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
      601 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
      661 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
      721 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
      781 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
      841 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
      901 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
      961 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1021 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     1081 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     1141 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     1201 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     1261 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     1321 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     1381 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     1441 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     1501 tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     1561 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     1621 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     1681 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     1741 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     1801 aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     1861 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     1921 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     1981 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     2041 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     2101 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     2161 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     2221 taggcgtatc acgaggccct ttcgtgttca agaattagct tggccattgc atacgttgta
     2281 tccatatcat aatatgtaca tttatattgg ctcatgtcca acattaccgc catgttgaca
     2341 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     2401 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     2461 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     2521 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     2581 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     2641 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     2701 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     2761 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     2821 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     2881 cggtaggcgt gtacggtggg aggtctatat aagcagagct caataaaaga gcccacaacc
     2941 cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tattcccaat
     3001 aaagcctctt gctgtttgca tccgaatcgt ggtctcgctg ttccttggga gggtctcctc
     3061 tgagtgattg actacccacg acgggggtct ttcatttggg ggctcgtccg ggatttggag
     3121 acccctgccc agggaccacc gacccaccac cgggaggtaa gctggccagc aacttatctg
     3181 tgtctgtccg attgtctagt gtctatgttt gatgttatgc gcctgcgtct gtactagtta
     3241 gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg aacacccggc
     3301 cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc gacctgagga
     3361 agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg agacgagaac
     3421 ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg aagccgcgcg
     3481 tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg
     3541 tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac cttaggtcac
     3601 tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa gagacgttgg
     3661 ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga gacggcacct
     3721 ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc ccgcatggac
     3781 acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac ccccctccct
     3841 gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc gccccgtctc
     3901 tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca gccctcactc
     3961 cttctctagg cgccggaatt gaagatcttt aattaacgat ctatcgattc tagatctaga
     4021 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
     4081 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
     4141 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
     4201 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
     4261 cgaaacaccg tagctggcat ttatgtaggg ttttagagct agaaatagca agttaaaata
     4321 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tgaatttaga
     4381 tcttgagaca aggtgccggg ccgggaattc ggcgaattgg gcccgacgtc gcatgctccc
     4441 ggcccctcga gtggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc
     4501 gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta
     4561 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
     4621 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
     4681 caggtgtcgt gacgcggcca ccatgaaccc agccatcagc gtcgctctcc tgctctcagt
     4741 cttgcaggtg tcccgagggc agaaggtgac cagcctgaca gcctgcctgg tgaaccaaaa
     4801 ccttcgcctg gactgccgcc atgagaataa caccaaggat aactccatcc agcatgagtt
     4861 cagcctgacc cgagagaaga ggaagcacgt gctctcaggc accctcggga tacccgagca
     4921 cacgtaccgc tcccgcgtca ccctctccaa ccagccctat atcaaggtcc ttaccctagc
     4981 caacttcacc accaaggatg agggcgacta cttttgtgag cttcgagtct cgggcgcgaa
     5041 tcccatgagc tccaataaaa gtatcagtgt gtatagagac aaactggtca agtgtggcgg
     5101 cataagcctg ctggttcaga acacatcctg gatgctgctg ctgctgcttt ccctctccct
     5161 cctccaagcc ctggacttca tttctctggg cagcggcgcc accaacttca gcctgctgaa
     5221 gcaggccggc gacgtggagg agaaccccgg ccccatgatt gaacaagatg gattgcacgc
     5281 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     5341 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     5401 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     5461 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     5521 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     5581 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     5641 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5701 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5761 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5821 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5881 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5941 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     6001 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgat tagatatcat
     6061 cagtggtcca ggctctagtt ttgactcaac aatatcacca gctgaagcct atagagtacg
     6121 agccatagat aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc
     6181 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata
     6241 cataactgag aatagagaag ttcagatcaa ggtcaggaac agatggaaca gggtcgaccc
     6301 tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat
     6361 ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc
     6421 aataaaagag cccacaaccc ctcactcggg gcgccagtcc tccgattgac tgagtcgccc
     6481 gggtacccgt gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt
     6541 ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt tcatttgggg
     6601 gctcgtccgg gatcgggaga cccctgccca gggaccaccg acccaccacc gggaggtaag
     6661 ctggctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     6721 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     6781 cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag
     6841 tgtagatccg gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     6901 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     6961 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     7021 tcccgcccct aactccgccc atcccgcccc taactc
//
