LOCUS       sequence-263323-        6984 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of ALK3 WT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6984)
  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 6984)
  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 wt
FEATURES             Location/Qualifiers
     source          1..6984
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        117..496
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        497..700
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     651..671
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     745..764
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        745..763
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2434..2460
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2440..2466
                     /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(2491..2509)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2492..2509)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2529..2546)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2535..2759
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2805..3233
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2892..2911)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3102..3123
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3242..3262)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3286..3496
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3308..3504
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3355..3490
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3417..3436
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3571..4365
                     /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(3625..3644)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4235..4254
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4539..4660
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4576..4595)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4630..4649
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4709..4725)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4709..4725)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4722..4744)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4733..4749
                     /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(4757..4763,4764..4781,4782..4787))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4802..4823
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4940..4957)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5111..5699)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5191..5210)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5870..6661,6662..6730))
                     /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     6493..6512
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6731..6835)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6910..6929)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
       61 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      121 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      181 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      241 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      301 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      361 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      421 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      481 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      541 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      601 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      661 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      721 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagcttggta
      781 ccgagctcgg atccactagt aacggccgcc agtgtgctgg aattcgccat ggctcagcta
      841 tacatttaca tcagattatt gggagcctat ttgttcatca tttctcgtgt tcaaggacag
      901 aatctggata gtatgcttca tggcactggg atgaaatcag actccgacca gaaaaagtca
      961 gaaaatggag taaccttagc accagaggat accttgcctt ttttaaagtg ctattgctca
     1021 gggcactgtc cagatgatgc tattaataac acatgcataa ctaatggaca ttgctttgcc
     1081 atcatagaag aagatgacca gggagaaacc acattagctt cagggtgtat gaaatatgaa
     1141 ggatctgatt ttcagtgcaa agattctcca aaagcccagc tacgccggac aatagaatgt
     1201 tgtcggacca atttatgtaa ccagtatttg caacccacac tgccccctgt tgtcataggt
     1261 ccgttttttg atggcagcat tcgatggctg gttttgctca tttctatggc tgtctgcata
     1321 attgctatga tcatcttctc cagctgcttt tgttacaaac attattgcaa gagcatctca
     1381 agcagacgtc gttacaatcg tgatttggaa caggatgaag catttattcc agttggagaa
     1441 tcactaaaag accttattga ccagtcacaa agttctggta gtgggtctgg actaccttta
     1501 ttggttcagc gaactattgc caaacagatt cagatggtcc ggcaagttgg taaaggccga
     1561 tatggagaag tatggatggg caaatggcgt ggcgaaaaag tggcggtgaa agtattcttt
     1621 accactgaag aagccagctg gtttcgagaa acagaaatct accaaactgt gctaatgcgc
     1681 catgaaaaca tacttggttt catagcggca gacattaaag gtacaggttc ctggactcag
     1741 ctctatttga ttactgatta ccatgaaaat ggatctctct atgacttcct gaaatgtgct
     1801 acactggaca ccagagccct gcttaaattg gcttattcag ctgcctgtgg tctgtgccac
     1861 ctgcacacag aaatttatgg cacccaagga aagcccgcaa ttgctcatcg agacctaaag
     1921 agcaaaaaca tcctcatcaa gaaaaatggg agttgctgca ttgctgacct gggccttgct
     1981 gttaaattca acagtgacac aaatgaagtt gatgtgccct tgaataccag ggtgggcacc
     2041 aaacgctaca tggctcccga agtgctggac gaaagcctga acaaaaacca cttccagccc
     2101 tacatcatgg ctgacatcta cagcttcggc ctaatcattt gggagatggc tcgtcgttgt
     2161 atcacaggag ggatcgtgga agaataccaa ttgccatatt acaacatggt accgagtgat
     2221 ccgtcatacg aagatatgcg tgaggttgtg tgtgtcaaac gtttgcggcc aattgtgtct
     2281 aatcggtgga acagtgatga atgtctacga gcagttttga agctaatgtc agaatgctgg
     2341 gcccacaatc cagcctccag actcacagca ttgagaatta agaagacgct tgccaagatg
     2401 gttgaatccc aagatgtaaa aatcctcgag ggttatccgt atgatgttcc tgattatgct
     2461 agcctctagg tcgactctag agggccctat tctatagtgt cacctaaatg ctagagctcg
     2521 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     2581 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     2641 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     2701 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     2761 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     2821 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     2881 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     2941 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3001 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3061 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3121 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3181 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     3241 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3301 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3361 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3421 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     3481 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     3541 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3601 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3661 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3721 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     3781 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3841 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3901 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3961 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4021 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4081 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4141 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4201 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4261 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4321 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4381 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4441 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4501 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     4561 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4621 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     4681 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4741 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4801 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4861 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4921 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4981 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5041 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5101 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5161 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5221 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5281 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5341 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5401 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5461 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5521 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5581 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5641 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5701 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5761 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5821 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5881 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5941 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6001 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6061 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6121 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6181 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6241 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6301 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6361 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6421 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6481 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6541 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6601 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6661 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6721 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6781 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6841 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg
     6901 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
     6961 tgtgttggag gtcgctgagt agtg
//
