LOCUS       sequence-157437-        7057 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of ALK3 K261R.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7057)
  AUTHORS   Raja E, Tzavlaki K, Vuilleumier R, Edlund K, Kahata K, Zieba A, 
            Moren A, Watanabe Y, Voytyuk I, Botling J, Soderberg O, Micke P, 
            Pyrowolakis G, Heldin CH, Moustakas A
  TITLE     The protein kinase LKB1 negatively regulates bone morphogenetic 
            protein receptor signaling.
  JOURNAL   Oncotarget. 2016 Jan 12;7(2):1120-43. doi: 10.18632/oncotarget.6683.
  PUBMED    26701726
REFERENCE   2  (bases 1 to 7057)
  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: 
            "10.18632/oncotarget"; journalName: "Oncotarget"; date: "2016-01-12-
            12"; volume: "7"; issue: "2"; pages: "1120-43"
COMMENT     Sequence Label: pcDNA3-ALK3 K261R
FEATURES             Location/Qualifiers
     source          1..7057
                     /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             2552..2578
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2558..2584
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     promoter        complement(2609..2627)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2610..2627)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2647..2664)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2653..2877
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2923..3351
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3010..3029)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3220..3241
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3360..3380)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3365..3695
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3546..3681
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3608..3627
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3762..4556
                     /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(3816..3835)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4426..4445
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4730..4851
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4767..4786)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4821..4840
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4900..4916)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4900..4916)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4913..4935)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4924..4940
                     /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(4948..4954,4955..4972,4973..4978))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4993..5014
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5131..5148)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5302..5890)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5382..5401)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6061..6852,6853..6921))
                     /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     6684..6703
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6922..7026)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttcgccatgg ctcagctata
      961 catttacatc agattattgg gagcctattt gttcatcatt tctcgtgttc aaggacagaa
     1021 tctggatagt atgcttcatg gcactgggat gaaatcagac tccgaccaga aaaagtcaga
     1081 aaatggagta accttagcac cagaggatac cttgcctttt ttaaagtgct attgctcagg
     1141 gcactgtcca gatgatgcta ttaataacac atgcataact aatggacatt gctttgccat
     1201 catagaagaa gatgaccagg gagaaaccac attagcttca gggtgtatga aatatgaagg
     1261 atctgatttt cagtgcaaag attctccaaa agcccagcta cgccggacaa tagaatgttg
     1321 tcggaccaat ttatgtaacc agtatttgca acccacactg ccccctgttg tcataggtcc
     1381 gttttttgat ggcagcattc gatggctggt tttgctcatt tctatggctg tctgcataat
     1441 tgctatgatc atcttctcca gctgcttttg ttacaaacat tattgcaaga gcatctcaag
     1501 cagacgtcgt tacaatcgtg atttggaaca ggatgaagca tttattccag ttggagaatc
     1561 actaaaagac cttattgacc agtcacaaag ttctggtagt gggtctggac tacctttatt
     1621 ggttcagcga actattgcca aacagattca gatggtccgg caagttggta aaggccgata
     1681 tggagaagta tggatgggca aatggcgtgg cgaaaaagtg gcggtgagag tattctttac
     1741 cactgaagaa gccagctggt ttcgagaaac agaaatctac caaactgtgc taatgcgcca
     1801 tgaaaacata cttggtttca tagcggcaga cattaaaggt acaggttcct ggactcagct
     1861 ctatttgatt actgattacc atgaaaatgg atctctctat gacttcctga aatgtgctac
     1921 actggacacc agagccctgc ttaaattggc ttattcagct gcctgtggtc tgtgccacct
     1981 gcacacagaa atttatggca cccaaggaaa gcccgcaatt gctcatcgag acctaaagag
     2041 caaaaacatc ctcatcaaga aaaatgggag ttgctgcatt gctgacctgg gccttgctgt
     2101 taaattcaac agtgacacaa atgaagttga tgtgcccttg aataccaggg tgggcaccaa
     2161 acgctacatg gctcccgaag tgctggacga aagcctgaac aaaaaccact tccagcccta
     2221 catcatggct gacatctaca gcttcggcct aatcatttgg gagatggctc gtcgttgtat
     2281 cacaggaggg atcgtggaag aataccaatt gccatattac aacatggtac cgagtgatcc
     2341 gtcatacgaa gatatgcgtg aggttgtgtg tgtcaaacgt ttgcggccaa ttgtgtctaa
     2401 tcggtggaac agtgatgaat gtctacgagc agttttgaag ctaatgtcag aatgctgggc
     2461 ccacaatcca gcctccagac tcacagcatt gagaattaag aagacgcttg ccaagatggt
     2521 tgaatcccaa gatgtaaaaa tcctcgaggg ttatccgtat gatgttcctg attatgctag
     2581 cctctaggtc gactctagag ggccctattc tatagtgtca cctaaatgct agagctcgct
     2641 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     2701 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2761 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2821 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     2881 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     2941 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3001 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3061 gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg
     3121 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3181 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3241 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt
     3301 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     3361 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccaggcaggc agaagtatgc
     3421 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3481 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3541 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3601 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     3661 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     3721 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3781 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3841 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3901 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     3961 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4021 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4081 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4141 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4201 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4261 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4321 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4381 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4441 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4501 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4561 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4621 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4681 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     4741 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4801 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4861 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4921 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4981 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5041 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5101 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5161 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5221 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5281 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5341 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5401 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5461 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     5521 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5581 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5641 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5701 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     5761 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5821 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5881 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5941 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6001 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6061 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6121 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6181 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6241 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6301 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6361 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6421 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6481 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6541 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6601 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6661 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6721 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6781 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6841 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6901 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6961 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7021 tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
