LOCUS       sequence-424726-        6868 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of ALK7 K222R.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6868)
  AUTHORS   Moren A, Itoh S, Moustakas A, Dijke P, Heldin CH
  TITLE     Functional consequences of tumorigenic missense mutations in the 
            amino-terminal domain of Smad4.
  JOURNAL   Oncogene. 2000 Sep 7;19(38):4396-404.
  PUBMED    10980615
REFERENCE   2  (bases 1 to 6868)
  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: "Oncogene.";
            date: "2000-09-7"; volume: "19(38)"; pages: "4396-404"
COMMENT     Sequence Label: pcDNA3-ALK7 K222R
FEATURES             Location/Qualifiers
     source          1..6868
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        93..472
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        473..676
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     627..647
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     721..740
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        721..739
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2294..2320
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2300..2326
                     /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(2351..2369)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2352..2369)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2389..2406)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2395..2619
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2665..3093
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2752..2771)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2962..2983
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3102..3122)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3146..3356
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3168..3364
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3215..3350
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3277..3296
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3431..4225
                     /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(3485..3504)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4095..4114
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4399..4520
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4436..4455)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4490..4509
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4569..4585)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4569..4585)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4582..4604)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4593..4609
                     /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(4617..4623,4624..4641,4642..4647))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4662..4683
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4800..4817)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4971..5559)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5051..5070)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5730..6521,6522..6590))
                     /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     6353..6372
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6591..6695)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6770..6789)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
       61 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
      121 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      181 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      241 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      301 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      361 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      421 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      481 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      541 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      601 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      661 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat
      721 taatacgact cactataggg agacccaagc ttggtaccga gctcggatcc actagtaacg
      781 gccgccagtg tgctggaatt ccaccatgac cccagcaagc cgctccgcac tgagcctggc
      841 cctcctgctg gtggcactgg cctccgacct tgcggcagga ctgaagtgtg tgtgtctttt
      901 gtgtgattcc tcaaacttta cctgccaaac cgaaggagca tgctgggcct ctgtcatgct
      961 aaccaacggg aaagaacagg tgatcaaatc gtgcgtgtcc ctcccggaac taaatgctca
     1021 ggtcttctgt cacagttcca acaacgtgac caagaccgaa tgttgcttca cagacttctg
     1081 caacaacatc actctgcacc ttcccacagc atctccagat gcccctagac ttggccccac
     1141 agagctgaca gttgttatca ctgtacctgt ttgcctcctg tccatcgcag ccatgctaac
     1201 gatatgggcc tgccaggacc gccagtgcac atacaggaag accaagagac acaatgtgga
     1261 ggaaccactg gcagagtaca gccttgtcaa tgctggaaaa accctcaaag atctgattta
     1321 tgatgccact gcctcgggct caggatctgg cctgcctctt ttggttcaaa gaaccatcgc
     1381 aaggacaatt gtacttcaag aaatcgtagg aaaaggtcgg tttggggaag tgtggcacgg
     1441 aagatggtgt ggagaagatg tggctgtgcg catattctcc tccagagatg agagatcttg
     1501 gttccgtgag gcagaaattt atcagacggt aatgctgaga catgagaata ttctcggttt
     1561 catcgcggcc gacaacaaag ataatggaac ctggactcag ctttggcttg tgtcagagta
     1621 tcacgagcag ggctccttat atgactattt gaatagaagc atagtgaccg tggctggaat
     1681 ggtcaagttg gcgctttcaa tagcgagtgg tctggctcac ctacacatgg agatcgtggg
     1741 cactcaaggt aagcctgcta ttgctcaccg agatataaag tcaaagaata tcttagtcaa
     1801 aaagtgtgac acttgtgcca tagctgactt agggctggct gtgaaacatg attctatcat
     1861 gaacactata gatatacccc agaatcctaa agtgggaacc aagaggtata tggctcccga
     1921 aatgcttgat gatacaatga acgtcaacat ctttgagtcc ttcaagcgag ctgacatcta
     1981 ttcggtgggg ctggtttact gggaaatagc tcgaaggtgt tcagttggag gacttgttga
     2041 agagtaccag ttgccttatt atgacatggt gccttcagat ccttccatag aggaaatgag
     2101 gaaggtcgtt tgtgatcaga aactccgacc aaatctccca aaccagtggc aaagctgtga
     2161 ggcgctccgg gtcatgggaa gaataatgcg tgagtgctgg tatgccaacg gggcagctcg
     2221 cctgaccgcc ctgcgcgtga agaagaccat ttctcagctg tgtgtcaagg aagactgtaa
     2281 ggccctcgag ggttatccgt atgatgttcc tgattatgct agcctctagg tcgactctag
     2341 agggccctat tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc
     2401 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     2461 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     2521 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     2581 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     2641 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2701 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2761 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2821 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2881 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2941 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3001 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     3061 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     3121 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3181 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3241 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3301 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3361 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     3421 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     3481 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     3541 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     3601 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     3661 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     3721 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     3781 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     3841 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     3901 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     3961 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4021 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     4081 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     4141 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     4201 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     4261 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4321 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4381 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4441 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4501 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     4561 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4621 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4681 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4741 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4801 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4861 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4921 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4981 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5041 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5101 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5161 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5221 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5281 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5341 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5401 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5461 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5521 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5581 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5641 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5701 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5761 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5821 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5881 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5941 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6001 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6061 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6121 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6181 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6241 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6301 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6361 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6421 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6481 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6541 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6601 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6661 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6721 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
     6781 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     6841 agtgcgcgag caaaatttaa gctacaac
//
