LOCUS       sequence-407172-        7345 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human MPL-HA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7345)
  AUTHORS   Araki M, Yang Y, Masubuchi N, Hironaka Y, Takei H, Morishita S, 
            Mizukami Y, Kan S, Shirane S, Edahiro Y, Sunami Y, Ohsaka A, Komatsu
            N
  TITLE     Activation of the thrombopoietin receptor by mutant calreticulin in 
            CALR-mutant myeloproliferative neoplasms.
  JOURNAL   Blood. 2016 Jan 27. pii: blood-2015-09-671172.
  PUBMED    26817954
REFERENCE   2  (bases 1 to 7345)
  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: "Blood. 2016
            Jan 27. pii: blood-2015-09-671172."
COMMENT     Sequence Label: pcDNA3.1/MPL-HA
FEATURES             Location/Qualifiers
     source          1..7345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2869..2895
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2936..2953)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2942..3166
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3212..3640
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3299..3318)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3509..3530
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3649..3669)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3654..3983
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3834..3969
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3896..3915
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4050..4844
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4104..4123)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4714..4733
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5018..5139
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5055..5074)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5109..5128
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5188..5204)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5188..5204)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5201..5223)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5212..5228
                     /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)."
     promoter        complement(join(5236..5242,5243..5260,5261..5266))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5281..5302
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5419..5436)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5590..6178)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5670..5689)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6349..7140,7141..7209))
                     /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     6972..6991
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7210..7314)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatgccct cctgggccct cttcatggtc acctcctgcc tcctcctggc ccctcaaaac
     1021 ctggcccaag tcagcagcca agatgtctcc ttgctggcat cagactcaga gcccctgaag
     1081 tgtttctccc gaacatttga ggacctcact tgcttctggg atgaggaaga ggcagcgccc
     1141 agtgggacat accagctgct gtatgcctac ccgcgggaga agccccgtgc ttgccccctg
     1201 agttcccaga gcatgcccca ctttggaacc cgatacgtgt gccagtttcc agaccaggag
     1261 gaagtgcgtc tcttctttcc gctgcacctc tgggtgaaga atgtgttcct aaaccagact
     1321 cggactcagc gagtcctctt tgtggacagt gtaggcctgc cggctccccc cagtatcatc
     1381 aaggccatgg gtgggagcca gccaggggaa cttcagatca gctgggagga gccagctcca
     1441 gaaatcagtg atttcctgag gtacgaactc cgctatggcc ccagagatcc caagaactcc
     1501 actggtccca cggtcataca gctgattgcc acagaaacct gctgccctgc tctgcagagg
     1561 cctcactcag cctctgctct ggaccagtct ccatgtgctc agcccacaat gccctggcaa
     1621 gatggaccaa agcagacctc cccaagtaga gaagcttcag ctctgacagc agagggtgga
     1681 agctgcctca tctcaggact ccagcctggc aactcctact ggctgcagct gcgcagcgaa
     1741 cctgatggga tctccctcgg tggctcctgg ggatcctggt ccctccctgt gactgtggac
     1801 ctgcctggag atgcagtggc acttggactg caatgcttta ccttggacct gaagaatgtt
     1861 acctgtcaat ggcagcaaca ggaccatgct agctcccaag gcttcttcta ccacagcagg
     1921 gcacggtgct gccccagaga caggtacccc atctgggaga actgcgaaga ggaagagaaa
     1981 acaaatccag gactacagac cccacagttc tctcgctgcc acttcaagtc acgaaatgac
     2041 agcattattc acatccttgt ggaggtgacc acagccccgg gtactgttca cagctacctg
     2101 ggctcccctt tctggatcca ccaggctgtg cgcctcccca ccccaaactt gcactggagg
     2161 gagatctcca gtgggcatct ggaattggag tggcagcacc catcgtcctg ggcagcccaa
     2221 gagacctgtt atcaactccg atacacagga gaaggccatc aggactggaa ggtgctggag
     2281 ccgcctctcg gggcccgagg agggaccctg gagctgcgcc cgcgatctcg ctaccgttta
     2341 cagctgcgcg ccaggctcaa cggccccacc taccaaggtc cctggagctc gtggtcggac
     2401 ccaactaggg tggagaccgc caccgagacc gcctggatct ccttggtgac cgctctgcat
     2461 ctagtgctgg gcctcagcgc cgtcctgggc ctgctgctgc tgaggtggca gtttcctgca
     2521 cactacagga gactgaggca tgccctgtgg ccctcacttc cagacctgca ccgggtccta
     2581 ggccagtacc ttagggacac tgcagccctg agcccgccca aggccacagt ctcagatacc
     2641 tgtgaagaag tggaacccag cctccttgaa atcctcccca agtcctcaga gaggactcct
     2701 ttgcccctgt gttcctccca ggcccagatg gactaccgaa gattgcagcc ttcttgcctg
     2761 gggaccatgc ccctgtctgt gtgcccaccc atggctgagt cagggtcctg ctgtaccacc
     2821 cacattgcca accattccta cctaccacta agctattggc agcagcctta cccatacgat
     2881 gttccagatt acgcttgact cgagtctaga gggcccgttt aaacccgctg atcagcctcg
     2941 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3001 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3061 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     3121 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     3181 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3241 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3301 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3361 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3421 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3481 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3541 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     3601 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     3661 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3721 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3781 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3841 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3901 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     3961 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     4021 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4081 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4141 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4201 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     4261 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4321 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4381 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4441 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4501 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4561 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4621 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4681 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4741 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4801 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4861 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4921 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4981 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5041 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5101 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5161 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5221 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5281 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5341 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5401 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5461 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5521 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5581 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5641 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5701 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5761 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5821 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5881 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5941 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6001 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6061 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6121 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6181 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6241 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6301 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6361 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6421 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6481 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6541 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6601 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6661 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6721 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6781 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6841 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6901 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6961 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7021 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7081 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7141 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7201 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7261 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7321 tccccgaaaa gtgccacctg acgtc
//
