LOCUS       sequence-407194-        7360 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human MPL-V5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7360)
  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 7360)
  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-V5
FEATURES             Location/Qualifiers
     source          1..7360
                     /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..2910
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2951..2968)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2957..3181
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3227..3655
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3314..3333)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3524..3545
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3664..3684)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3669..3998
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3849..3984
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3911..3930
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4065..4859
                     /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(4119..4138)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4729..4748
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5033..5154
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5070..5089)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5124..5143
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5203..5219)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5203..5219)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5216..5238)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5227..5243
                     /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(5251..5257,5258..5275,5276..5281))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5296..5317
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5434..5451)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5605..6193)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5685..5704)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6364..7155,7156..7224))
                     /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     6987..7006
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7225..7329)
                     /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 agcagcctgg taagcctatc
     2881 cctaaccctc tcctcggtct cgattctacg tgactcgagt ctagagggcc cgtttaaacc
     2941 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3001 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3061 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3121 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     3181 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     3241 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3301 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3361 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3421 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3481 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3541 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3601 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     3661 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3721 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3781 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3841 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3901 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     3961 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     4021 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4081 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4141 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4201 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     4261 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4321 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4381 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4441 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4501 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4561 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     4621 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4681 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4741 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4801 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4861 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4921 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4981 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     5041 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5101 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     5161 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     5221 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5281 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5341 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5401 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5461 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5521 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5581 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5641 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5701 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5761 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5821 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5881 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5941 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6001 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6061 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6121 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6181 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6241 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6301 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6361 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6421 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6481 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6541 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6601 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6661 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6721 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6781 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6841 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6901 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6961 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7021 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7081 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7141 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7201 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7261 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7321 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
