LOCUS       sequence-364062-        9232 bp    DNA     circular SYN 15-APR-2025
DEFINITION  expressing mouse Arkadia (isoform 3) C946S H948L E3 ligase dead 
            mutant proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9232)
  AUTHORS   Xu H, Wu L, Nguyen HH, Mesa KR, Raghavan V, Episkopou V, Littman DR
  TITLE     Arkadia-SKI/SnoN signaling differentially regulates TGF-beta-induced
            iTreg and Th17 cell differentiation.
  JOURNAL   J Exp Med. 2021 Nov 1;218(11). pii: 212614. doi: 
            10.1084/jem.20210777. Epub 2021 Sep 2.
  PUBMED    34473197
REFERENCE   2  (bases 1 to 9232)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Exp 
            Med."; date: "2021-11-1"; volume: "218(11). pii: 212614. doi"; 
            pages: " 10.1084/jem.20210777. Epub 2021 Sep 2"
COMMENT     Sequence Label: MSCV-Arkadia C946S H948L
FEATURES             Location/Qualifiers
     source          1..9232
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    574..915
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             982..1398
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1328..1350
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1366..1382
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             2944..2964
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     protein_bind    complement(4402..4426)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    4460..5026
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4476..5028
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4620..4637)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4847..4866
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5076..5564
                     /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"
     LTR             5620..6134
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    6296..6312
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6320..6326,6327..6344,6345..6350))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6365..6386
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6503..6520)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6674..7262)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6754..6773)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7433..8224,8225..8293))
                     /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     8056..8075
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8294..8398)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8466..8484
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8522..8544)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8644..8663
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8857..8879
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     9007..9026
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(9226..9232,1..510)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa
       61 atacataact gagaatagag aagttcagat caaggttagg aacagagaga cagcagaata
      121 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga
      181 tggtccccag atgcggtccc gccctcagca gtttctagag aaccatcaga tgtttccagg
      241 gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc
      301 tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac
      361 tcggcgcgcc agtcctccga tagactgcgt cgcccgggta cccgtattcc caataaagcc
      421 tcttgctgtt tgcatccgaa tcgtggactc gctgatcctt gggagggtct cctcagattg
      481 attgactgcc cacctcgggg gtctttcatt tggaggttcc accgagattt ggagacccct
      541 gcccagggac caccgacccc cccgccggga ggtaagctgg ccagcggtcg tttcgtgtct
      601 gtctctgtct ttgtgcgtgt ttgtgccggc atctaatgtt tgcgcctgcg tctgtactag
      661 ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc
      721 ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga
      781 ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag
      841 aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc
      901 gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt
      961 ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt
     1021 cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt
     1081 tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc
     1141 acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat
     1201 ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct
     1261 ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg
     1321 tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc
     1381 actccttctc taggcgccgg aattagatcc atgtctcaat ggactcctga atttaacgag
     1441 ctctacaccc taaaagtggc tatgaagagt gggactcctg atgcgcccac tacacaggag
     1501 agtctgaagg cggtccttct gcacccacag cccctagggg ccaccaagag cttccctgca
     1561 gaagtggaga tgattaatag caaagtgggg aatgaattct ctcatctgtg tgatgattct
     1621 caaaaacaag agaaggacat gactggcaac cagcaagaac aagaaaaaag tggtgttgtg
     1681 agaaaaaagc gcaaaagcca gcaggctggc ccctcatatg tgcagaactg tgtcaaagaa
     1741 aatcaggaaa tcctagggcg cagacaacag ctagagacac cgagtgatga ggataatgac
     1801 tcttccttaa gtgagtgtct gtcttctccc tcatctagtc tccattttgg gggctctgat
     1861 acagtgactt cagatgagga taaagaagtc tcggtgagac atacccagcc tgttctgagt
     1921 gctaaaagta gaagtcacag tgcaaggtct cacaagtggc ctcgaactga ggcagatcct
     1981 gtgccaagtt tgttaatgaa aagaccttgt tttcatggca gtgcactaag gagagtcaca
     2041 tgcagaaaga ggctggtgaa gagcagctcc tcgcagagga cacagaagca gaaggagagg
     2101 atgctagtgc agaggaagaa gcgggaggcg ctagctcaga ggaagtatgc tcttctctcc
     2161 agctccagta gctccagtga gaatgacctc agcagcgatt cctcgtccag ctcctccacc
     2221 gatggagaag aggacctgtg tgcatctgcc agtgagaacc ccagcaaccc cgctgctccc
     2281 tcaggaagta ttgatgaaga tgttgtggtg atagaagcct ccttcactcc ccaggtcact
     2341 gccaatgaag aaattaatgt tacctcaact gacagtgaag tggagattgt tacagttgga
     2401 gaaagctatc ggtctcgttc aacccttgga cactccagat ctcattggag ccaaggttca
     2461 agttcgcata caggacggcc acaggagtct cgaaaccgca gtaggatctc tactgttata
     2521 cagcccctga ggcagaatgc agcagaagtt gtggacctta ctgttgatga agatgaacct
     2581 actatagtac caaccacttc tgctagaatg gactcacaaa ctactagtgc ttccattaac
     2641 aattcaaatc catctacctc tgagcaagcc tctgatacta cttcaactgt ggccagtagc
     2701 cagccttcca cagtgtcaga gactgaagct actcttacaa gcaatagtgc aactggttct
     2761 tctgtgggca atgacgtgag gagaactgca tctagtgctg tcccagaaag tggtcctcct
     2821 gcaatgccaa ggctaccttc ttgctgcccc cagcactcac catgtggagg gacatcacag
     2881 agccatcatg cattagcaca cccacactca agttgttttc agcagcatgg ccaccatttt
     2941 cagcatcacc accatcacca ccatactccc cacccagctg tcccagtttc tccttccttc
     3001 agtgaccctg cttgcccagt ggaaagacct caagtacagg cgccttgtgg agcaaatagc
     3061 agctctggtt ccagctacca tgaccaggca ttgccagtcg atctgagcaa cagtgctctc
     3121 aggactcatg gaagtggggg tttccatggt gcatctgctt ttgacccctg ctgccctgtc
     3181 acatcttccc gggctgcagt ctttggccat caggctgctg ctgccccgac tcagccatta
     3241 tcaatagatg gctatggctc cagcatggtt gcccagcccc agccccagcc ccctccacag
     3301 ccttctcttt cgtcatgtcg gcactacatg ccacctccgt atgcttcctt gacaagacca
     3361 ctgcaccatc aagcctcagc ttgccatcac tctcatggaa atgcccctcc ccagactcag
     3421 cctccccctc aagtggatta cgtgattccg catcctgtcc atgctttcca ttctcagata
     3481 tcttctcatg cagcatctca ccctgtggca ccaccgccgc caacacattt aggcagtaca
     3541 gcggcaccca tccctcagca tcttcctcct gctcaccagc cgatttcaca ccatattcca
     3601 gccccagcac cttcagccca gagactgcac cctcatgaag tgatgcagag gatggaagtt
     3661 cagaggagga ggatgatgca gcatccaacg cgggcacatg aacggccccc gccccatcca
     3721 cataggatgc acccaaacta tggccatggg catcatattc atgtgcctca aaccatgtct
     3781 tcacatcctc gccaggctcc agagagaact gcctgggaac tggggattga agctggagta
     3841 actgcagcta cctacacacc tggtgctctg caccctcacc tggcccatta ccacgccccg
     3901 cctcgacttc atcacctcca gttaggagca cttcctttaa tggttcctga catggcaggc
     3961 tatcctcaca tccgttatat ttcttcagga ttggatggag catcattcag aggtcctttt
     4021 aggggcaatt ttgaggaatt gattcatctg gaagaaagat taggaaatgt caatcgtgga
     4081 gcctcccagg gaacaattga aagatgcaca tatccacata aatataaaaa gaggaaactg
     4141 cactgcaaac aagatgggga agaagggact gaggaagaca cagaggaaaa atgtactatc
     4201 tgtttgtcta ttttagagga aggtgaagat gtgaggcgcc ttccatcaat gttgcttttc
     4261 caccaagtgt gtgttgatca gtggctgatc accaataaga agtgccccat atgcagagtg
     4321 gacatcgagg cccagctgcc aagtgaaagt tgagtcgact ggatccggta ccgaattcgc
     4381 ggccgcactc gagatatcta gacccagctt tcttgtacaa agtggttcga tgacggtatc
     4441 gataagcttg atatcgaatt ccgccccccc ccctaacgtt actggccgaa gccgcttgga
     4501 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa
     4561 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
     4621 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc
     4681 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
     4741 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca
     4801 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag
     4861 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct
     4921 ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc
     4981 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg gccacaacca
     5041 tgggtctttt ctgcagtcac cgtcctcgag gcaccatgaa cccagccatc agcgtcgctc
     5101 tcctgctctc agtcttgcag gtgtcccgag ggcagaaggt gaccagcctg acagcctgcc
     5161 tggtgaacca aaaccttcgc ctggactgcc gccatgagaa taacaccaag gataactcca
     5221 tccagcatga gttcagcctg acccgagaga agaggaagca cgtgctctca ggcacccttg
     5281 ggatacccga gcacacgtac cgctcccgcg tcaccctctc caaccagccc tatatcaagg
     5341 tccttaccct agccaacttc accaccaagg atgagggcga ctacttttgt gagcttcgcg
     5401 tctcgggcgc gaatcccatg agctccaata aaagtatcag tgtgtataga gacaagctgg
     5461 tcaagtgtgg cggcataagc ctgctggttc agaacacatc ctggatgctg ctgctgctgc
     5521 tttccctctc cctcctccaa gccctggact tcatttctct gtgacgcgta gaattcgata
     5581 aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg
     5641 tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaataca taactgagaa
     5701 tagagaagtt cagatcaagg ttaggaacag agagacagca gaatatgggc caaacaggat
     5761 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
     5821 gtcccgccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
     5881 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
     5941 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
     6001 tccgatagac tgcgtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
     6061 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
     6121 cgggggtctt tcatgggtaa cagtttcttg aagttggaga acaacattct gagggtagga
     6181 gtcgaatatt aagtaatcct gactcaatta gccactgttt tgaatccaca tactccaata
     6241 ctcctgaaat agttcattat ggacagcgca gaaagagctg gggagaattg tgaaattgtt
     6301 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     6361 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6421 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6481 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6541 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6601 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6661 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6721 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6781 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6841 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6901 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6961 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7021 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7081 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     7141 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7201 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7261 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7321 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7381 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7441 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7501 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7561 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7621 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7681 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7741 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7801 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7861 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7921 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7981 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     8041 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8101 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8161 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8221 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8281 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8341 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8401 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     8461 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     8521 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     8581 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     8641 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     8701 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
     8761 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     8821 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     8881 cggcgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc
     8941 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
     9001 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
     9061 ggccacgatg cgtccggcgt agaggcgatt agtccaattt gttaaagaca ggatatcagt
     9121 ggtccaggct ctagttttga ctcaacaata tcaccagctg aagcctatag agtacgagcc
     9181 atagataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga aa
//
