LOCUS       sequence-456745-        7077 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of ALK6 K231R.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7077)
  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 7077)
  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-ALK6 K231R
FEATURES             Location/Qualifiers
     source          1..7077
                     /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             2645..2671
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2651..2677
                     /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(2702..2720)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2703..2720)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2740..2757)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2746..2970
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3016..3444
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3103..3122)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3313..3334
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3453..3473)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3497..3707
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3519..3715
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3566..3701
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3628..3647
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3782..4576
                     /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(3836..3855)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4446..4465
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4750..4871
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4787..4806)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4841..4860
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4920..4936)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4920..4936)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4933..4955)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4944..4960
                     /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(4968..4974,4975..4992,4993..4998))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5013..5034
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5151..5168)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5322..5910)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5402..5421)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6081..6872,6873..6941))
                     /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     6704..6723
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6942..7046)
                     /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 gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttcaagcggc ggcagaagtt
      961 gccggcgtgg tgctcgtagt gagggcgcgg aggacccggg acctgggaag cggcggcggg
     1021 ttaacttcgg ctgaatcaca accatttggc gctgagctat gacaagagag caaacaaaaa
     1081 gttaaaggag caacccggcc ataagtgaag agagaagttt attgataaca tgctcttacg
     1141 aagctctgga aaattaaatg tgggcaccaa gaaggaggat ggagagagta cagcccccac
     1201 ccctcggccc aagatcctac gttgtaaatg ccaccaccac tgtccggaag actcagtcaa
     1261 caatatctgc agcacagatg ggtactgctt cacgatgata gaagaagatg actctggaat
     1321 gcctgttgtc acctctggat gtctaggact agaagggtca gattttcaat gtcgtgacac
     1381 tcccattcct catcaaagaa gatcaattga atgctgcaca gaaaggaatg agtgtaataa
     1441 agacctccac cccactctgc ctcctctcaa ggacagagat tttgttgatg ggcccataca
     1501 ccacaaggcc ttgcttatct ctgtgactgt ctgtagttta ctcttggtcc tcattatttt
     1561 attctgttac ttcaggtata aaagacaaga agcccgacct cggtacagca ttgggctgga
     1621 gcaggacgag acatacattc ctcctggaga gtccctgaga gacttgatcg agcagtctca
     1681 gagctcggga agtggatcag gcctccctct gctggtccaa aggacaatag ctaagcaaat
     1741 tcagatggtg aagcagattg gaaaaggccg ctatggcgag gtgtggatgg gaaagtggcg
     1801 tggagaaaag gtggctgtga gagtgttctt caccacggag gaagccagct ggttccgaga
     1861 gactgagata tatcagacgg tcctgatgcg gcatgagaat attctggggt tcattgctgc
     1921 agatatcaaa gggactgggt cctggactca gttgtacctc atcacagact atcatgaaaa
     1981 cggctccctt tatgactatc tgaaatccac caccttagac gcaaagtcca tgctgaagct
     2041 agcctactcc tctgtcagcg gcctatgcca tttacacacg gaaatcttta gcactcaagg
     2101 caagccagca atcgcccatc gagacttgaa aagtaaaaac atcctggtga agaaaaatgg
     2161 aacttgctgc atagcagacc tgggcttggc tgtcaagttc attagtgaca caaatgaggt
     2221 tgacatccca cccaacaccc gggttggcac caagcgctat atgcctccag aagtgctgga
     2281 cgagagcttg aatagaaacc atttccagtc ctacattatg gctgacatgt acagctttgg
     2341 actcatcctc tgggagattg caaggagatg tgtttctgga ggtatagtgg aagaatacca
     2401 gcttccctat cacgacctgg tgcccagtga cccttcttat gaggacatga gagaaattgt
     2461 gtgcatgaag aagttacggc cttcattccc caatcgatgg agcagtgatg agtgtctcag
     2521 gcagatgggg aagcttatga cagagtgctg ggcgcagaat cctgcctcca ggctgacggc
     2581 cctgagagtt aagaaaaccc ttgccaaaat gtcagagtcc caggacatta aactcctcga
     2641 gggttatccg tatgatgttc ctgattatgc tagcctctag gtcgactcta gagggcccta
     2701 ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg ccttctagtt
     2761 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     2821 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     2881 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     2941 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     3001 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3061 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3121 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3181 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3241 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3301 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3361 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3421 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     3481 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3541 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3601 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     3661 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3721 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     3781 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3841 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3901 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3961 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4021 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4081 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4141 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4201 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4261 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     4321 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4381 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4441 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4501 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4561 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4621 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4681 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4741 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4801 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4861 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     4921 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4981 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5041 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5101 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5161 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5221 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5281 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5341 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5401 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5461 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5521 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5581 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5641 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5701 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5761 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5821 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5881 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5941 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6001 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6061 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6121 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6181 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6241 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6301 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6361 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6421 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6481 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6541 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6601 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6661 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6721 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6781 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6841 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6901 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6961 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7021 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
