LOCUS       sequence-259540-        5813 bp    DNA     circular SYN 14-APR-2025
DEFINITION  expresses HA-human PP2A C-alpha D88N in Hi5 insect cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5813)
  AUTHORS   Cho US, Xu W
  TITLE     Crystal structure of a protein phosphatase 2A heterotrimeric 
            holoenzyme.
  JOURNAL   Nature. 2007 Jan 4;445(7123):53-7. doi: 10.1038/nature05351. Epub 
            2006 Nov 1.
  PUBMED    17086192
REFERENCE   2  (bases 1 to 5813)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nature05351"; journalName: "Nature"; date: "2007-01-4- 4"; 
            volume: "445"; issue: "7123"; pages: "53-7"
COMMENT     Sequence Label: pFastbacHTB HA-human PP2A C-alpha D88N
FEATURES             Location/Qualifiers
     source          1..5813
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..18
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             40..60
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             76..102
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1242..1376
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1279..1298)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1333..1352
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  1405..1570
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      1754..2208
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1841..1860)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2050..2071
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2234..2338
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2339..2407,2408..3199)
                     /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(2557..2576)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3370..3958
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3859..3878
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4112..4129
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  4263..4487
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(4554..5087)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(5276..5281,5282..5298,5299..5304))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        5656..5747
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     5672..5689
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
ORIGIN
        1 catcaccatc accatcacga ttacgatatc ccaacgaccg aaaacctgta ttttcagggc
       61 gccatgggat ccatgtaccc atatgatgtt ccagattatg ctgacgagaa ggtgttcacc
      121 aaggagctgg accagtggat cgagcagctg aacgagtgca agcagctgtc cgagtcccag
      181 gtcaagagcc tctgcgagaa ggctaaagaa atcctgacaa aagaatccaa cgtgcaagag
      241 gttcgatgtc cagttactgt ctgtggagat gtgcatgggc aatttcatga tctcatggaa
      301 ctgtttagaa ttggtggcaa atcaccagat acaaattact tgtttatggg agattatgtt
      361 aacagaggat attattcagt tgaaacagtt acactgcttg tagctcttaa ggttcgttac
      421 cgtgaacgca tcaccattct tcgagggaat catgagagca gacagatcac acaagtttat
      481 ggtttctatg atgaatgttt aagaaaatat ggaaatgcaa atgtttggaa atattttaca
      541 gatctttttg actatcttcc tctcactgcc ttggtggatg ggcagatctt ctgtctacat
      601 ggtggtctct cgccatctat agatacactg gatcatatca gagcacttga tcgcctacaa
      661 gaagttcccc atgagggtcc aatgtgtgac ttgctgtggt cagatccaga tgaccgtggt
      721 ggttggggta tatctcctcg aggagctggt tacacctttg ggcaagatat ttctgagaca
      781 tttaatcatg ccaatggcct cacgttggtg tctagagctc accagctagt gatggaggga
      841 tataactggt gccatgaccg gaatgtagta acgattttca gtgctccaaa ctattgttat
      901 cgttgtggta accaagctgc aatcatggaa cttgacgata ctctaaaata ctctttcttg
      961 cagtttgacc cagcacctcg tagaggcgag ccacatgtta ctcgtcgtac cccagactac
     1021 ttcctgtaag aattcaaagg cctacgtcga cgagctcact agtcgcggcc gctttcgaat
     1081 ctagagcctg cagtctcgag gcatgcggta ccaagcttgt cgagaagtac tagaggatca
     1141 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     1201 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     1261 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     1321 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatctgat
     1381 cactgatatc gcctaggaga tccgaaccag ataagtgaaa tctagttcca aactattttg
     1441 tcatttttaa ttttcgtatt agcttacgac gctacaccca gttcccatct attttgtcac
     1501 tcttccctaa ataatcctta aaaactccat ttccacccct cccagttccc aactattttg
     1561 tccgcccaca gcggggcatt tttcttcctg ttatgttttt aatcaaacat cctgccaact
     1621 ccatgtgaca aaccgtcatc ttcggctact ttttctctgt cacagaatga aaatttttct
     1681 gtcatctctt cgttattaat gtttgtaatt gactgaatat caacgcttat ttgcagcctg
     1741 aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     1801 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     1861 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     1921 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     1981 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     2041 ttcttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     2101 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     2161 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
     2221 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     2281 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     2341 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     2401 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     2461 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     2521 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     2581 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     2641 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     2701 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     2761 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     2821 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     2881 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     2941 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     3001 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     3061 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     3121 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     3181 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     3241 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     3301 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3361 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3421 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3481 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     3541 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3601 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3661 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3721 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     3781 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     3841 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3901 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     3961 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     4021 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     4081 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     4141 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagacca
     4201 gccgcgtaac ctggcaaaat cggttacggt tgagtaataa atggatgccc tgcgtaagcg
     4261 ggtgtgggcg gacaataaag tcttaaactg aacaaaatag atctaaacta tgacaataaa
     4321 gtcttaaact agacagaata gttgtaaact gaaatcagtc cagttatgct gtgaaaaagc
     4381 atactggact tttgttatgg ctaaagcaaa ctcttcattt tctgaagtgc aaattgcccg
     4441 tcgtattaaa gaggggcgtg gccaagggca tggtaaagac tatattcgcg gcgttgtgac
     4501 aatttaccga acaactccgc ggccgggaag ccgatctcgg cttgaacgaa ttgttaggtg
     4561 gcggtacttg ggtcgatatc aaagtgcatc acttcttccc gtatgcccaa ctttgtatag
     4621 agagccactg cgggatcgtc accgtaatct gcttgcacgt agatcacata agcaccaagc
     4681 gcgttggcct catgcttgag gagattgatg agcgcggtgg caatgccctg cctccggtgc
     4741 tcgccggaga ctgcgagatc atagatatag atctcactac gcggctgctc aaacttgggc
     4801 agaacgtaag ccgcgagagc gccaacaacc gcttcttggt cgaaggcagc aagcgcgatg
     4861 aatgtcttac tacggagcaa gttcccgagg taatcggagt ccggctgatg ttgggagtag
     4921 gtggctacgt ctccgaactc acgaccgaaa agatcaagag cagcccgcat ggatttgact
     4981 tggtcagggc cgagcctaca tgtgcgaatg atgcccatac ttgagccacc taactttgtt
     5041 ttagggcgac tgccctgctg cgtaacatcg ttgctgctgc gtaacatcgt tgctgctcca
     5101 taacatcaaa catcgaccca cggcgtaacg cgcttgctgc ttggatgccc gaggcataga
     5161 ctgtacaaaa aaacagtcat aacaagccat gaaaaccgcc actgcgccgt taccaccgct
     5221 gcgttcggtc aaggttctgg accagttgcg tgagcgcata cgctacttgc attacagttt
     5281 acgaaccgaa caggcttatg tcaactgggt tcgtgccttc atccgtttcc acggtgtgcg
     5341 tcacccggca accttgggca gcagcgaagt cgaggcattt ctgtcctggc tggcgaacga
     5401 gcgcaaggtt tcggtctcca cgcatcgtca ggcattggcg gccttgctgt tcttctacgg
     5461 caaggtgctg tgcacggatc tgccctggct tcaggagatc ggaagacctc ggccgtcgcg
     5521 gcgcttgccg gtggtgctga ccccggatga agtggttcgc atcctcggtt ttctggaagg
     5581 cgagcatcgt ttgttcgccc aggactctag ctatagttct agtggttggc tacgtatact
     5641 ccggaatatt aatagatcat ggagataatt aaaatgataa ccatctcgca aataaataag
     5701 tattttactg ttttcgtaac agttttgtaa taaaaaaacc tataaatatt ccggattatt
     5761 cataccgtcc caccatcggg cgcggatctc ggtccgaaac catgtcgtac tac
//
