LOCUS       sequence-356779-        6274 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of 10*His tagged human SHP1 phosphatase domain 
            (243-541aa).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6274)
  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 6274)
  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: pET28a-His10-SHP1_PTPase (243-541aa)
FEATURES             Location/Qualifiers
     source          1..6274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      58..513
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(145..164)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     355..376
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(605..1420)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(605..1420)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1328..1347
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1412..1431
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1542..2130
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2031..2050
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2284..2301
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2316..2455
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2381..2400)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2541..2563
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2557..2748)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3557..4639)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4601..4620
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4640..4717)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4923..4942
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5026..5045
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5026..5044
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5045..5069
                     /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)."
     RBS             5084..5106
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5125..5154
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             5164..5181
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             5185..5217
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             6164..6181
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6234..6252)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      join(6248..6274,1..21)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc gaatgggacg
       61 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
      121 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
      181 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
      241 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
      301 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
      361 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
      421 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
      481 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
      541 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta
      601 attcttagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
      661 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
      721 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
      781 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
      841 cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag
      901 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
      961 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     1021 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     1081 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     1141 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     1201 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     1261 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     1321 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     1381 gcggcctaga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     1441 ttatgtaagc agacagtttt attgttcatg accaaaatcc cttaacgtga gttttcgttc
     1501 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     1561 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     1621 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     1681 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     1741 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     1801 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     1861 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     1921 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     1981 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     2041 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     2101 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     2161 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     2221 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     2281 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     2341 catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg
     2401 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
     2461 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     2521 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     2581 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     2641 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     2701 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt
     2761 gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac
     2821 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
     2881 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt
     2941 taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa
     3001 cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa
     3061 gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg
     3121 ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt
     3181 cctcaacgac aggagcacga tcatgcgcac ccgtggggcc gccatgccgg cgataatggc
     3241 ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg
     3301 caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc
     3361 ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa
     3421 gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg
     3481 gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac
     3541 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     3601 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt
     3661 ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga
     3721 gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg
     3781 ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat
     3841 ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat
     3901 cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt
     3961 gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc
     4021 gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc
     4081 ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg
     4141 taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa
     4201 ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg
     4261 gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac
     4321 aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg
     4381 cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg
     4441 caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt
     4501 aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg
     4561 cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt
     4621 ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg
     4681 ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta
     4741 tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc
     4801 gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
     4861 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
     4921 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
     4981 acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaattaata cgactcacta
     5041 taggggaatt gtgagcggat aacaattccc ctctagaaat aattttgttt aactttaaga
     5101 aggagatata ccatgggcag cagccatcat catcatcatc atcatcacca tcacagcagc
     5161 ggcctggtgc cgcgcggcag ccatatggct agcatgactg gtggacagca aatgggtcgc
     5221 ggaggcttct gggaggagtt tgagagtttg cagaagcagg aggtgaagaa cttgcaccag
     5281 cgtctggaag ggcagcggcc agagaacaag ggcaagaacc gctacaagaa cattctcccc
     5341 tttgaccaca gccgagtgat cctgcaggga cgggacagta acatccccgg gtccgactac
     5401 atcaatgcca actacatcaa gaaccagctg ctaggccctg atgagaacgc taagacctac
     5461 atcgccagcc agggctgtct ggaggccacg gtcaatgact tctggcagat ggcgtggcag
     5521 gagaacagcc gtgtcatcgt catgaccacc cgagaggtgg agaaaggccg gaacaaatgc
     5581 gtcccatact ggcccgaggt gggcatgcag cgtgcttatg ggccctactc tgtgaccaac
     5641 tgcggggagc atgacacaac cgaatacaaa ctccgtacct tacaggtctc cccgctggac
     5701 aatggagacc tgattcggga gatctggcat taccagtacc tgagctggcc cgaccatggg
     5761 gtccccagtg agcctggggg tgtcctcagc ttcctggacc agatcaacca gcggcaggaa
     5821 agtctgcctc acgcagggcc catcatcgtg cactgcagcg ccggcatcgg ccgcacaggc
     5881 accatcattg tcatcgacat gctcatggag aacatctcca ccaagggcct ggactgtgac
     5941 attgacatcc agaagaccat ccagatggtg cgggcgcagc gctcgggcat ggtgcagacg
     6001 gaggcgcagt acaagttcat ctacgtggcc atcgcccagt tcattgaaac cactaagaag
     6061 aagctggagg tcctgcagtc gcagaagggc caggagtcgg agtacgggaa catcacctat
     6121 taagaattcg agctccgtcg acaagcttgc ggccgcactc gagcaccacc accaccacca
     6181 ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     6241 gcaataacta gcataacccc ttggggcctc taaa
//
