LOCUS       sequence-356813-        7793 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mouse SHP2 fused to N-terminal mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7793)
  AUTHORS   Xu X, Hou B, Fulzele A, Masubuchi T, Zhao Y, Wu Z, Hu Y, Jiang Y, Ma
            Y, Wang H, Bennett EJ, Fu G, Hui E
  TITLE     PD-1 and BTLA regulate T cell signaling differentially and only 
            partially through SHP1 and SHP2.
  JOURNAL   J Cell Biol. 2020 Jun 1;219(6). pii: 151801. doi: 
            10.1083/jcb.201905085.
  PUBMED    32437509
REFERENCE   2  (bases 1 to 7793)
  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"; journalName: "J Cell 
            Biol."; date: "2020-06-1"; volume: "219(6). pii: 151801. doi"; 
            pages: " 10.1083/jcb.201905085"
COMMENT     Sequence Label: pMIG II SIN-SV40-mCherry-SHP2
FEATURES             Location/Qualifiers
     source          1..7793
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     37..56
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     290..307
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    424..445
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(460..465,466..483,484..490)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    498..514
                     /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)."
     CDS             complement(3009..3716)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3284..3303)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3553..3571
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     promoter        complement(3736..3946)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3742..3877
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             complement(3973..4389)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(3989..4005)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(4021..4043)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    4456..4797
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             4861..5377
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(5577..5596)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(5724..5746)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5940..5959)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6059..6081
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6119..6137)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6205..6309
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6310..6378,6379..7170)
                     /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     complement(6528..6547)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7341..7793,1..136)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
       61 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
      121 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
      181 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
      241 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
      301 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
      361 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
      421 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
      481 tcgtatgttg tgtggaattg tgagcggata acaatttcac aattctcccc agctctttct
      541 gcgctgtcca taatgaacta tttcaggagt attggagtat gtggattcaa aacagtggct
      601 aattgagtca ggattactta atattcgact cctaccctca gaatgttgtt ctccaacttc
      661 aagaaactgt tacccatgaa agacccccgc tgacgggtag tcaatcactc agaggagacc
      721 ctcccaagga acagcgagac cacaagtcgg atgcaactgc aagagggttt attggataca
      781 cgggtacccg ggcgacgcag tctatcggag gactggcgcg ccgagtgagg ggttgtgggc
      841 tcttttattg agctcgggga gcagaagcgc gcgaacagaa gcgagaagcg aactgattgg
      901 ttagttcaaa taaggcacag ggtcatttca ggtccttggg gcaccctgga aacatctgat
      961 ggttcttgcc aaacctacag gtggggtctt tcattccccc ctttttctgg agactaaata
     1021 aaatctttta ttttatcgat atcagcgctt taaatttgcg catgctagct atagttctag
     1081 aggtaccggt tgttaacgtt agccggctac ctgaagcttg catgcctgca ggtcgactct
     1141 agagtcgcgg ccgctcatct gaaactcctc tgctgctgca tgaggcccac gttctcatag
     1201 actcgggcgc tgtcctccct catttctgca cagggtggcg ttggggtgca gggtggcagg
     1261 ggactctgat caccacttgt ctggtccacc aaggaatact taatattggt atattcatgt
     1321 ccttttcttt tgcttttctg ctcctcctcg atccggcgct gcagcgtctc tatgtagtgc
     1381 tggacagcca tgtagataaa ccggtactgt gcttctgtct ggaccatccc cgacctctgg
     1441 gaccgcacca tctgaatggt tttaggaacg tcgatgtcac agtccacacc tttctctcga
     1501 atgatgtcaa taaggatgtc aatcacaatg aaggttcctg tccggccaat cccagcgctg
     1561 cagtgaacca cgacagggcc tgcatccacg atgctctcct gcttgtggtg gacctcctcc
     1621 aggaagtcca gcacacctcc agggtcacta ggcacgccat ggtctggcca ggtccgaaag
     1681 tggtactgcc agacggttct ctctgtgttt ccctggagta gagcttgtcc gaccttagag
     1741 agtttgagct ctcgtaaagt gtagtcatgg gcggcacttt ctttgacgtt cctaacacgc
     1801 atgaccccgt attctttgag cgcatactca tcaggccagt acttgacaca tttgctcttc
     1861 cctctctcca cttcctttgt ggtcatgaca atgactcgag agttctcctg gaacaccatc
     1921 cgccagaagt cattcaccgt gttttgcagg cagccttgag tggcaatgta actctttttg
     1981 ggtttggaat tgttgcactt ggtctcaaac tcaggcatga tgatgtttgc attaatgtaa
     2041 tcagaaacag gctcattggg atccccatca tgcagaacga ccctggtatg atcaaagggc
     2101 aggatgtttt tgtatctgtt tttgttttta ttttcttgtc tctgtccttc ttttcggcta
     2161 tagagaagtt tgcattcctg ttgctggagc gtctcaaact cttcccaaaa gccctgcttg
     2221 actttatctg tggtctcagc cagcttgctt aactctcgaa cccggctttc aatttcagca
     2281 gcattgatac gagttgtgtt gaggggctgc ttgagctgca ggactgtgcc cagcgtctcc
     2341 accatggggt tcttcttgta atgctccacc aggtctgtca gagagtcaaa gcgctctccc
     2401 ccaccaacgt cgtatttcag ctcctgacag cggatcatga cgtgggtcac tttggacttg
     2461 ccgtcgttgc tctccccttt gtcgtcacca gtgcgcacgg agagaacgaa gtctccgggg
     2521 tggctctggc tctctcgaac gaggaagctg ccatgcttgc ccttctccgt cagcagcttc
     2581 tctgcttctt ttccagacaa gtgaccatgg aaccaccttt cagaggtagg gtctgcacag
     2641 ttcagcgggt acttgagctc gataacatct ccattcttct ctttcagctg cccatggtgt
     2701 tccatgtaat actgaaccag ttcagccaaa gtggcaaact tctccccacc atagaggtca
     2761 tagtagtccc cagtgttctg aatcttgatg tgggtaacag ctccatttct tctaacagac
     2821 agagtgaagt ctccagggtt actcttactg ggccttgcta aaaaactgcc atcgactcct
     2881 ctggtcagca ggagattctc tgcctccaca ccagtgatgt tggggtgaaa ccatctccgc
     2941 gatgtcatgc ggccgccacc gctcgagcct ccacctccgc tcccgccacc accggaactc
     3001 ccacgcgtct tgtacagctc gtccatgccg ccggtggagt ggcggccctc ggcgcgttcg
     3061 tactgttcca cgatggtgta gtcctcgttg tgggaggtga tgtccaactt gatgttgacg
     3121 ttgtaggcgc cgggcagctg cacgggcttc ttggccttgt aggtggtctt gacctcagcg
     3181 tcgtagtggc cgccgtcctt cagcttcagc ctctgcttga tctcgccctt cagggcgccg
     3241 tcctcggggt acatccgctc ggaggaggcc tcccagccca tggtcttctt ctgcattacg
     3301 gggccgtcgg aggggaagtt ggtgccgcgc agcttcacct tgtagatgaa ctcgccgtcc
     3361 tgcagggagg agtcctgggt cacggtcacc acgccgccgt cctcgaagtt catcacgcgc
     3421 tcccacttga agccctcggg gaaggacagc ttcaagtagt cggggatgtc ggcggggtgc
     3481 ttcacgtagg ccttggagcc gtacatgaac tgaggggaca ggatgtccca ggcgaagggc
     3541 agggggccac ccttggtcac cttcagcttg gcggtctggg tgccctcgta ggggcggccc
     3601 tcgccctcgc cctcgatctc gaactcgtgg ccgttcacgg agccctccat gtgcaccttg
     3661 aagcgcatga actccttgat gatggccatg ttatcctcct cgcccttgct caccatacgc
     3721 gtgagaattc tcgagagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct
     3781 cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg
     3841 gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta
     3901 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgcctct cgagagctcc
     3961 agatcgaatt ccggcgccta gagaaggagt gagggctgga taaagggagg atcgaggcgg
     4021 ggtcgaacga ggaggttcaa gggggagaga cggggcggat ggaggaagag gaggcggagg
     4081 cttagggtgt acaaagggct tgacccaggg aggggggtca aaagccaagg cttcccaggt
     4141 cacgatgtag gggacctggt ctgggtgtcc atgcgggcca ggtgaaaaga ccttgatctt
     4201 aacctgggtg atgaggtctc ggttaaaggt gccgtctcgc ggccatccga cgttaaaggt
     4261 tggccattct gcagagcaga aggtaaccca acgtctcttc ttgacatcta ccgactggtt
     4321 gtgagcgatc cgctcgacat ctttccagtg acctaaggtc aaacttaagg gagtggtaac
     4381 agtctggccc taattttcag acaaatacag aaacacagtc agacagagac aacacagaac
     4441 gatgctgcag cgctgcagca gacaagacgc gcggcttcgg ttccaaaccg aaagcaaaaa
     4501 ttcagacgga ggcgggaact gttttaggtt ctcgtctcct accagaacca catatcctga
     4561 cggggtcgga ttccacatcg actcccttcc tcaggtcggg ccacaaaaac ggcccccaaa
     4621 gtccctggga cgtctcccag ggttgcggcc gggtgttcag aactcgtcag ttccaccacg
     4681 ggtccgccag atacagagct agttagctaa ctagtacaga cgcaggcgca aacattagat
     4741 gccggcacaa acacgcacaa agacagagac agacacgaaa cgaccgctgg ccagcttacc
     4801 tcccggcggg ggggtcggtg gtccctgggc aggggtctcc aaatctcggt ggaacctcca
     4861 aatgaaagac ccccgaggtg ggcagtcaat caatctgagg agaccctccc aaggatcagc
     4921 gagtccacga ttcggatgca aacagcaaga ggctttattg ggaatacggg tacccgggcg
     4981 acgcagtcta tcggaggact ggcgcgccga gtgaggggtt gtgggctctt ttattgagct
     5041 cggggagcag aagcgcgcga acagaagcga gaagcgaact gattggttag ttcaaataag
     5101 gcacagggtc atttcaggtc cttggggcac cctggaaaca tctgatggtt ctctagaaac
     5161 tgctgagggc gggaccgcat ctggggacca tctgttcttg gccctgagcc ggggcaggaa
     5221 ctgcttacca cagatatcct gtttggccca tattctgctg tctctctgtt cctaaccttg
     5281 atctgaactt ctctattctc agttatgtat tttccatgcc ttgcaaaatg gcgttactta
     5341 agctagcttg ccaaacctac aggtggggtc tttcattccc ccctttttct ggagactaaa
     5401 taaaatcttt tattttatct atggctcgta ctctataggc ttcagctggt gatattgttg
     5461 agtcaaaact agagcctgga ccactgatat cctgtcttta acaaattgga ctaatcgcct
     5521 ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg ctggcgccta
     5581 tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca tgagcgcttg
     5641 tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg ccatctcctt
     5701 gcatgcacca ttccttgcgc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     5761 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     5821 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg
     5881 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
     5941 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     6001 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     6061 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     6121 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     6181 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     6241 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     6301 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     6361 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     6421 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     6481 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     6541 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     6601 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     6661 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6721 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6781 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6841 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     6901 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     6961 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     7021 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     7081 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     7141 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     7201 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     7261 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     7321 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7381 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7441 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     7501 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7561 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7621 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7681 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7741 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cag
//
