LOCUS       sequence-242321-        6944 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6944)
  AUTHORS   O'Reilly AM, Pluskey S, Shoelson SE, Neel BG
  TITLE     Activated mutants of SHP-2 preferentially induce elongation of 
            Xenopus animal caps.
  JOURNAL   Mol Cell Biol 2000 Jan;20(1):299-311.
  PUBMED    10594032
REFERENCE   2  (bases 1 to 6944)
  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: "Mol Cell 
            Biol"; date: "2000-01"; volume: "20(1)"; pages: "299-311"
COMMENT     Sequence Label: pGEX-4T1 SHP2 WT
FEATURES             Location/Qualifiers
     source          1..6944
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(178..183,184..199,200..206)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    214..230
                     /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             253..906
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     864..886
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             913..930
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2989..3011)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3156..3174)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3242..3346
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3347..3415,3416..4207)
                     /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     complement(3565..3584)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4378..4966
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4867..4886
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5120..5137
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5210..5287
                     /gene="lacI"
                     /label=lacI promoter
     CDS             5288..6370
                     /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     complement(5307..5326)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6419..6424,6425..6442,6443..6449)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6457..6473
                     /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)."
     primer_bind     6481..6497
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6481..6497
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(6493..6944,1..76)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(6513..6530)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6513..6529)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6522..6544)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 atcgactgca cggtgcacca atgcttctgg cgtcaggcag ccatcggaag ctgtggtatg
       61 gctgtgcagg tcgtaaatca ctgcataatt cgtgtcgctc aaggcgcact cccgttctgg
      121 ataatgtttt ttgcgccgac atcataacgg ttctggcaaa tattctgaaa tgagctgttg
      181 acaattaatc atcggctcgt ataatgtgtg gaattgtgag cggataacaa tttcacacag
      241 gaaacagtat tcatgtcccc tatactaggt tattggaaaa ttaagggcct tgtgcaaccc
      301 actcgacttc ttttggaata tcttgaagaa aaatatgaag agcatttgta tgagcgcgat
      361 gaaggtgata aatggcgaaa caaaaagttt gaattgggtt tggagtttcc caatcttcct
      421 tattatattg atggtgatgt taaattaaca cagtctatgg ccatcatacg ttatatagct
      481 gacaagcaca acatgttggg tggttgtcca aaagagcgtg cagagatttc aatgcttgaa
      541 ggagcggttt tggatattag atacggtgtt tcgagaattg catatagtaa agactttgaa
      601 actctcaaag ttgattttct tagcaagcta cctgaaatgc tgaaaatgtt cgaagatcgt
      661 ttatgtcata aaacatattt aaatggtgat catgtaaccc atcctgactt catgttgtat
      721 gacgctcttg atgttgtttt atacatggac ccaatgtgcc tggatgcgtt cccaaaatta
      781 gtttgtttta aaaaacgtat tgaagctatc ccacaaattg ataagtactt gaaatccagc
      841 aagtatatag catggccttt gcagggctgg caagccacgt ttggtggtgg cgaccatcct
      901 ccaaaatcgg atctggttcc gcgtggatcc ccaggaattc cgggcgggag gaacatgaca
      961 tcgcggagat ggtttcaccc aaatatcact ggtgtggagg cagaaaacct actgttgaca
     1021 agaggagttg atggcagttt tttggcaagg cctagtaaaa gtaaccctgg agacttcaca
     1081 ctttccgtta gaagaaatgg agctgtcacc cacatcaaga ttcagaacac tggtgattac
     1141 tatgacctgt atggagggga gaaatttgcc actttggctg agttggtcca gtattacatg
     1201 gaacatcacg ggcaattaaa agagaagaat ggagatgtca ttgagcttaa atatcctctg
     1261 aactgtgcag atcctacctc tgaaaggtgg tttcatggac atctctctgg gaaagaagca
     1321 gagaaattat taactgaaaa aggaaaacat ggtagttttc ttgtacgaga gagccagagc
     1381 caccctggag attttgttct ttctgtgcgc actggtgatg acaaagggga gagcaatgac
     1441 ggcaagtcta aagtgaccca tgttatgatt cgctgtcagg aactgaaata cgacgttggt
     1501 ggaggagaac ggtttgattc tttgacagat cttgtggaac attataagaa gaatcctatg
     1561 gtggaaacat tgggtacagt actacaactc aagcagcccc ttaacacgac tcgtataaat
     1621 gctgctgaaa tagaaagcag agttcgagaa ctaagcaaat tagctgagac cacagataaa
     1681 gtcaaacaag gcttttggga agaatttgag acactacaac aacaggagtg caaacttctc
     1741 tacagccgaa aagagggtca aaggcaagaa aacaaaaaca aaaatagata taaaaacatc
     1801 ctgccctttg atcataccag ggttgtccta cacgatggtg atcccaatga gcctgtttca
     1861 gattacatca atgcaaatat catcatgcct gaatttgaaa ccaagtgcaa caattcaaag
     1921 cccaaaaaga gttacattgc cacacaaggc tgcctgcaaa acacggtgaa tgacttttgg
     1981 cggatggtgt tccaagaaaa ctcccgagtg attgtcatga caacgaaaga agtggagaga
     2041 ggaaagagta aatgtgtcaa atactggcct gatgagtatg ctctaaaaga atatggcgtc
     2101 atgcgtgtta ggaacgtcaa agaaagcgcc gctcatgact atacgctaag agaacttaaa
     2161 ctttcaaagg ttggacaagg gaatacggag agaacggtct ggcaatacca ctttcggacc
     2221 tggccggacc acggcgtgcc cagcgaccct gggggcgtgc tggacttcct ggaggaggtg
     2281 caccataagc aggagagcat catggatgca gggccggtcg tggtgcactg cagtgctgga
     2341 attggccgga cagggacgtt cattgtgatt gatattctta ttgacatcat cagagagaaa
     2401 ggtgttgact gcgatattga cgttcccaaa accatccaga tggtgcggtc tcagaggtca
     2461 gggatggtcc agacagaagc acagtaccga tttatctata tggcggtcca gcattatatt
     2521 gaaacactac agcgcaggat tgaagaagag cagaaaagca agaggaaagg gcacgaatat
     2581 acaaatatta agtattctct agcggaccag acgagtggag atcagagccc tctcccgcct
     2641 tgtactccaa cgccaccctg tgcagaaatg agagaagaca gtgctagagt ctatgaaaac
     2701 gtgggcctga tgcaacagca gaaaagtttc agatgagaaa acctgccaaa acttcagcac
     2761 agaaatagat gtggactttc accctctccc taaaaagatc aagaacagac gcaagaaagt
     2821 ttatgtgaag acagaatttg gatttggaag gcttgcaatg tggttgacta ccttttgata
     2881 agcaaaattt gaaaccattt aaagaccgga attcccgggt cgactcgagc ggccgcatcg
     2941 tgactgactg acgatctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     3001 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     3061 tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag
     3121 cgatagcgga gtgtataatt cttgaagacg aaagggcctc gtgatacgcc tatttttata
     3181 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     3241 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3301 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     3361 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     3421 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     3481 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     3541 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     3601 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     3661 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     3721 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     3781 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     3841 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc
     3901 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     3961 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     4021 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     4081 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     4141 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     4201 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     4261 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4321 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4381 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4441 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4501 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     4561 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4621 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4681 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4741 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4801 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4861 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4921 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4981 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     5041 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     5101 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     5161 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc ataaattccg acaccatcga
     5221 atggcgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc aattcagggt
     5281 ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg tctcttatca
     5341 gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc gggaaaaagt
     5401 ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac aactggcggg
     5461 caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg cgccgtcgca
     5521 aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat
     5581 ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg
     5641 cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg ctgtggaagc
     5701 tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac ccatcaacag
     5761 tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg tcgcattggg
     5821 tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc gtctgcgtct
     5881 ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg
     5941 cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt
     6001 tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac
     6061 cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg ataccgaaga
     6121 cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc tgctggggca
     6181 aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct
     6241 gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc aaaccgcctc
     6301 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     6361 cgggcagtga gcgcaacgca attaatgtaa gttagctcac tcattaggca ccccaggctt
     6421 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     6481 caggaaacag ctatgaccat gattacggat tcactggccg tcgttttaca acgtcgtgac
     6541 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     6601 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     6661 ggcgaatggc gctttgcctg gtttccggca ccagaagcgg tgccggaaag ctggctggag
     6721 tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa actggcagat gcacggttac
     6781 gatgcgccca tctacaccaa cgtgacctat cccattacgg tcaatccgcc gtttgttccc
     6841 acggagaatc cgacgggttg ttactcgctc acatttaatg ttgatgaaag ctggctacag
     6901 gaaggccaga cgcgaattat ttttgatggc gttggaatta gctt
//
