LOCUS       sequence-353468-        7880 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Baculo expression of GST tag fused to SNAP tag and human Shp2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7880)
  AUTHORS   Hui E, Cheung J, Zhu J, Su X, Taylor MJ, Wallweber HA, Sasmal DK, 
            Huang J, Kim JM, Mellman I, Vale RD
  TITLE     T cell costimulatory receptor CD28 is a primary target for 
            PD-1-mediated inhibition.
  JOURNAL   Science. 2017 Mar 31;355(6332):1428-1433. doi: 
            10.1126/science.aaf1292. Epub 2017 Mar 9.
  PUBMED    28280247
REFERENCE   2  (bases 1 to 7880)
  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: 
            "10.1126/science.aaf1292"; journalName: "Science"; date: 
            "2017-03-31- 31"; volume: "355"; issue: "6332"; pages: "1428-1433"
COMMENT     Sequence Label: pFastBac (GST-SNAP-Shp2)
FEATURES             Location/Qualifiers
     source          1..7880
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(89..108)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        482..586
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(587..655,656..1447)
                     /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(805..824)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1618..2206
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2107..2126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2360..2377
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2511..2735
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2802..3335)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3505..3510,3511..3527,3528..3533))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        3885..3976
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3901..3918
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4064..4084
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             4133..4786
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     4744..4766
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             4793..4816
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4823..5368
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="optimized for bacterial expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHEIKLLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYQQLAALAGNPAATAAVKTALSGNPVPILIPCHRVVSSSGAVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     polyA_signal    7370..7504
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7407..7426)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7461..7480
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  7533..7698
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
       61 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      181 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt cttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
      481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
      541 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
      601 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
      661 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
      721 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
      781 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
      841 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
      901 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
      961 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1021 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1081 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     1141 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     1201 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     1261 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     1321 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     1381 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     1441 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     1501 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     1561 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1621 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     1681 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     1741 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     1801 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     1861 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     1921 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     1981 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2041 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2101 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2161 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2221 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2281 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2341 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     2401 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct
     2461 ggcaaaatcg gttacggttg agtaataaat ggatgccctg cgtaagcggg tgtgggcgga
     2521 caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt cttaaactag
     2581 acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat actggacttt
     2641 tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc gtattaaaga
     2701 ggggcgtggc caagggcatg gtaaagacta tattcgcggc gttgtgacaa tttaccgaac
     2761 aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg
     2821 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg
     2881 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca
     2941 tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact
     3001 gcgagatcat agatatagat ctcactacgc ggctgctcaa acttgggcag aacgtaagcc
     3061 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta
     3121 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct
     3181 ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg
     3241 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg
     3301 ccctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac ggcgtaacgc
     3361 gcttgctgct tggatgcccg aggcatagac tgtacaaaaa aacagtcata acaagccatg
     3421 aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt
     3481 gagcgcatac gctacttgca ttacagttta cgaaccgaac aggcttatgt caactgggtt
     3541 cgtgccttca tccgtttcca cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc
     3601 gaggcatttc tgtcctggct ggcgaacgag cgcaaggttt cggtctccac gcatcgtcag
     3661 gcattggcgg ccttgctgtt cttctacggc aaggtgctgt gcacggatct gccctggctt
     3721 caggagatcg gaagacctcg gccgtcgcgg cgcttgccgg tggtgctgac cccggatgaa
     3781 gtggttcgca tcctcggttt tctggaaggc gagcatcgtt tgttcgccca ggactctagc
     3841 tatagttcta gtggttggct acgtatactc cggaatatta atagatcatg gagataatta
     3901 aaatgataac catctcgcaa ataaataagt attttactgt tttcgtaaca gttttgtaat
     3961 aaaaaaacct ataaatattc cggattattc ataccgtccc accatcgggc gcggatctcg
     4021 gtccgaaacc atgtcgtact acgattacga tatcccaacg accgaaaacc tgtattttca
     4081 gggcgccatg ggatccggaa ttcaaaggcc tacgtcgacg agctcaacta gaatgtcccc
     4141 tatactaggt tattggaaaa ttaagggcct tgtgcaaccc actcgacttc ttttggaata
     4201 tcttgaagaa aaatatgaag agcatttgta tgagcgcgat gaaggtgata aatggcgaaa
     4261 caaaaagttt gaattgggtt tggagtttcc caatcttcct tattatattg atggtgatgt
     4321 taaattaaca cagtctatgg ccatcatacg ttatatagct gacaagcaca acatgttggg
     4381 tggttgtcca aaagagcgtg cagagatttc aatgcttgaa ggagcggttt tggatattag
     4441 atacggtgtt tcgagaattg catatagtaa agactttgaa actctcaaag ttgattttct
     4501 tagcaagcta cctgaaatgc tgaaaatgtt cgaagatcgt ttatgtcata aaacatattt
     4561 aaatggtgat catgtaaccc atcctgactt catgttgtat gacgctcttg atgttgtttt
     4621 atacatggac ccaatgtgcc tggatgcgtt cccaaaatta gtttgtttta aaaaacgtat
     4681 tgaagctatc ccacaaattg ataagtactt gaaatccagc aagtatatag catggccttt
     4741 gcagggctgg caagccacgt ttggtggtgg cgaccatcct ccaaaatcgg atctggaagt
     4801 tctgttccag gggcccacta gaatggacaa agattgcgaa atgaaacgta ccaccctgga
     4861 tagcccgctg ggcaaactgg aactgagcgg ctgcgaacag ggcctgcatg aaattaaact
     4921 gctgggtaaa ggcaccagcg cggccgatgc ggttgaagtt ccggccccgg ccgccgtgct
     4981 gggtggtccg gaaccgctga tgcaggcgac cgcgtggctg aacgcgtatt ttcatcagcc
     5041 ggaagcgatt gaagaatttc cggttccggc gctgcatcat ccggtgtttc agcaggagag
     5101 ctttacccgt caggtgctgt ggaaactgct gaaagtggtt aaatttggcg aagtgattag
     5161 ctatcagcag ctggcggccc tggcgggtaa tccggcggcc accgccgccg ttaaaaccgc
     5221 gctgagcggt aacccggtgc cgattctgat tccgtgccat cgtgtggtta gctctagcgg
     5281 tgcggttggc ggttatgaag gtggtctggc ggtgaaagag tggctgctgg cccatgaagg
     5341 tcatcgtctg ggtaaaccgg gtctgggagg tagcggtagc ggtagcggtg gaggtggaag
     5401 cagcatgaca tcgcggagat ggtttcaccc aaatatcact ggtgtggagg cagaaaacct
     5461 actgttgaca agaggagttg atggcagttt tttggcaagg cctagtaaaa gtaaccctgg
     5521 agacttcaca ctttccgtta gaagaaatgg agctgtcacc cacatcaaga ttcagaacac
     5581 tggtgattac tatgacctgt atggagggga gaaatttgcc actttggctg agttggtcca
     5641 gtattacatg gaacatcacg ggcaattaaa agagaagaat ggagatgtca ttgagcttaa
     5701 atatcctctg aactgtgcag atcctacctc tgaaaggtgg tttcatggac atctctctgg
     5761 gaaagaagca gagaaattat taactgaaaa aggaaaacat ggtagttttc ttgtacgaga
     5821 gagccagagc caccctggag attttgttct ttctgtgcgc actggtgatg acaaagggga
     5881 gagcaatgac ggcaagtcta aagtgaccca tgttatgatt cgctgtcagg aactgaaata
     5941 cgacgttggt ggaggagaac ggtttgattc tttgacagat cttgtggaac attataagaa
     6001 gaatcctatg gtggaaacat tgggtacagt actacaactc aagcagcccc ttaacacgac
     6061 tcgtataaat gctgctgaaa tagaaagcag agttcgagaa ctaagcaaat tagctgagac
     6121 cacagataaa gtcaaacaag gcttttggga agaatttgag acactacaac aacaggagtg
     6181 caaacttctc tacagccgaa aagagggtca aaggcaagaa aacaaaaaca aaaatagata
     6241 taaaaacatc ctgccctttg atcataccag ggttgtccta cacgatggtg atcccaatga
     6301 gcctgtttca gattacatca atgcaaatat catcatgcct gaatttgaaa ccaagtgcaa
     6361 caattcaaag cccaaaaaga gttacattgc cacacaaggc tgcctgcaaa acacggtgaa
     6421 tgacttttgg cggatggtgt tccaagaaaa ctcccgagtg attgtcatga caacgaaaga
     6481 agtggagaga ggaaagagta aatgtgtcaa atactggcct gatgagtatg ctctaaaaga
     6541 atatggcgtc atgcgtgtta ggaacgtcaa agaaagcgcc gctcatgact atacgctaag
     6601 agaacttaaa ctttcaaagg ttggacaagg gaatacggag agaacggtct ggcaatacca
     6661 ctttcggacc tggccggacc acggcgtgcc cagcgaccct gggggcgtgc tggacttcct
     6721 ggaggaggtg caccataagc aggagagcat catggatgca gggccggtcg tggtgcactg
     6781 cagtgctgga attggccgga cagggacgtt cattgtgatt gatattctta ttgacatcat
     6841 cagagagaaa ggtgttgact gcgatattga cgttcccaaa accatccaga tggtgcggtc
     6901 tcagaggtca gggatggtcc agacagaagc acagtaccga tttatctata tggcggtcca
     6961 gcattatatt gaaacactac agcgcaggat tgaagaagag cagaaaagca agaggaaagg
     7021 gcacgaatat acaaatatta agtattctct agcggaccag acgagtggag atcagagccc
     7081 tctcccgcct tgtactccaa cgccaccctg tgcagaaatg agagaagaca gtgctagagt
     7141 ctatgaaaac gtgggcctga tgcaacagca gaaaagtttc agatgaacta gtgcggccgc
     7201 tttcgaatct agagcctgca gtctcgaggc atgcggtacc aagcttgtcg agaagtacta
     7261 gaggatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     7321 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     7381 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     7441 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     7501 gatctgatca ctgcttgagc ctaggagatc cgaaccagat aagtgaaatc tagttccaaa
     7561 ctattttgtc atttttaatt ttcgtattag cttacgacgc tacacccagt tcccatctat
     7621 tttgtcactc ttccctaaat aatccttaaa aactccattt ccacccctcc cagttcccaa
     7681 ctattttgtc cgcccacagc ggggcatttt tcttcctgtt atgtttttaa tcaaacatcc
     7741 tgccaactcc atgtgacaaa ccgtcatctt cggctacttt ttctctgtca cagaatgaaa
     7801 atttttctgt catctcttcg ttattaatgt ttgtaattga ctgaatatca acgcttattt
     7861 gcagcctgaa tggcgaatgg
//
