LOCUS       sequence-353386-        7206 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Baculo expression of His10 tag fused to human Shp2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7206)
  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 7206)
  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(His10-Shp2)
FEATURES             Location/Qualifiers
     source          1..7206
                     /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             4043..4072
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             4094..4114
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             4118..4663
                     /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    6696..6830
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6733..6752)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6787..6806
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  6859..7024
                     /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 gcctgatgtg
     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 accatcacca tcaccatcac catcaccatc acgattacga
     4081 tatcccaacg accgaaaacc tgtattttca gggcgccatg gacaaagatt gcgaaatgaa
     4141 acgtaccacc ctggatagcc cgctgggcaa actggaactg agcggctgcg aacagggcct
     4201 gcatgaaatt aaactgctgg gtaaaggcac cagcgcggcc gatgcggttg aagttccggc
     4261 cccggccgcc gtgctgggtg gtccggaacc gctgatgcag gcgaccgcgt ggctgaacgc
     4321 gtattttcat cagccggaag cgattgaaga atttccggtt ccggcgctgc atcatccggt
     4381 gtttcagcag gagagcttta cccgtcaggt gctgtggaaa ctgctgaaag tggttaaatt
     4441 tggcgaagtg attagctatc agcagctggc ggccctggcg ggtaatccgg cggccaccgc
     4501 cgccgttaaa accgcgctga gcggtaaccc ggtgccgatt ctgattccgt gccatcgtgt
     4561 ggttagctct agcggtgcgg ttggcggtta tgaaggtggt ctggcggtga aagagtggct
     4621 gctggcccat gaaggtcatc gtctgggtaa accgggtctg ggaggtagcg gtagcggtag
     4681 cggtggaggt ggaagcagca tgacatcgcg gagatggttt cacccaaata tcactggtgt
     4741 ggaggcagaa aacctactgt tgacaagagg agttgatggc agttttttgg caaggcctag
     4801 taaaagtaac cctggagact tcacactttc cgttagaaga aatggagctg tcacccacat
     4861 caagattcag aacactggtg attactatga cctgtatgga ggggagaaat ttgccacttt
     4921 ggctgagttg gtccagtatt acatggaaca tcacgggcaa ttaaaagaga agaatggaga
     4981 tgtcattgag cttaaatatc ctctgaactg tgcagatcct acctctgaaa ggtggtttca
     5041 tggacatctc tctgggaaag aagcagagaa attattaact gaaaaaggaa aacatggtag
     5101 ttttcttgta cgagagagcc agagccaccc tggagatttt gttctttctg tgcgcactgg
     5161 tgatgacaaa ggggagagca atgacggcaa gtctaaagtg acccatgtta tgattcgctg
     5221 tcaggaactg aaatacgacg ttggtggagg agaacggttt gattctttga cagatcttgt
     5281 ggaacattat aagaagaatc ctatggtgga aacattgggt acagtactac aactcaagca
     5341 gccccttaac acgactcgta taaatgctgc tgaaatagaa agcagagttc gagaactaag
     5401 caaattagct gagaccacag ataaagtcaa acaaggcttt tgggaggaat ttgagacact
     5461 acaacaacag gagtgcaaac ttctctacag ccgaaaagag ggtcaaaggc aagaaaacaa
     5521 aaacaaaaat agatataaaa acatcctgcc ctttgatcat accagggttg tcctacacga
     5581 tggtgatccc aatgagcctg tttcagatta catcaatgca aatatcatca tgcctgaatt
     5641 tgaaaccaag tgcaacaatt caaagcccaa aaagagttac attgccacac aaggctgcct
     5701 gcaaaacacg gtgaatgact tttggcggat ggtgttccaa gaaaactccc gagtgattgt
     5761 catgacaacg aaagaagtgg agagaggaaa gagtaaatgt gtcaaatact ggcctgatga
     5821 gtatgctcta aaagaatatg gcgtcatgcg tgttaggaac gtcaaagaaa gcgccgctca
     5881 tgactatacg ctaagagaac ttaaactttc aaaggttgga caagggaata cggagagaac
     5941 ggtctggcaa taccactttc ggacctggcc ggaccacggc gtgcccagcg accctggggg
     6001 cgtgctggac ttcctggagg aggtgcacca taagcaggag agcatcatgg atgcagggcc
     6061 ggtcgtggtg cactgcagtg ctggaattgg ccggacaggg acgttcattg tgattgatat
     6121 tcttattgac atcatcagag agaaaggtgt tgactgcgat attgacgttc ccaaaaccat
     6181 ccagatggtg cggtctcaga ggtcagggat ggtccagaca gaagcacagt accgatttat
     6241 ctatatggcg gtccagcatt atattgaaac actacagcgc aggattgaag aagagcagaa
     6301 aagcaagagg aaagggcacg aatatacaaa tattaagtat tctctagcgg accagacgag
     6361 tggagatcag agccctctcc cgccttgtac tccaacgcca ccctgtgcag aaatgagaga
     6421 agacagtgct agagtctatg aaaacgtggg cctgatgcaa cagcagaaaa gtttcagatg
     6481 acggaattca aaggcctacg tcgacgagct caactagtgc ggccgctttc gaatctagag
     6541 cctgcagtct cgaggcatgc ggtaccaagc ttgtcgagaa gtactagagg atcataatca
     6601 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     6661 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     6721 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6781 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc tgatcactgc
     6841 ttgagcctag gagatccgaa ccagataagt gaaatctagt tccaaactat tttgtcattt
     6901 ttaattttcg tattagctta cgacgctaca cccagttccc atctattttg tcactcttcc
     6961 ctaaataatc cttaaaaact ccatttccac ccctcccagt tcccaactat tttgtccgcc
     7021 cacagcgggg catttttctt cctgttatgt ttttaatcaa acatcctgcc aactccatgt
     7081 gacaaaccgt catcttcggc tactttttct ctgtcacaga atgaaaattt ttctgtcatc
     7141 tcttcgttat taatgtttgt aattgactga atatcaacgc ttatttgcag cctgaatggc
     7201 gaatgg
//
