LOCUS       sequence-157381-        6976 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of ALK4 T206D.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6976)
  AUTHORS   ten Dijke P, Yamashita H, Ichijo H, Franzen P, Laiho M, Miyazono K, 
            Heldin CH
  TITLE     Characterization of type I receptors for transforming growth 
            factor-beta and activin.
  JOURNAL   Science. 1994 Apr 1;264(5155):101-4.
  PUBMED    8140412
REFERENCE   2  (bases 1 to 6976)
  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: "Science."; 
            date: "1994-04-1"; volume: "264(5155)"; pages: "101-4"
COMMENT     Sequence Label: pcDNA3 ALK4 T206D
FEATURES             Location/Qualifiers
     source          1..6976
                     /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             2471..2497
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2477..2503
                     /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(2528..2546)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2529..2546)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2566..2583)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2572..2796
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2842..3270
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2929..2948)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3139..3160
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3279..3299)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3284..3614
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3465..3600
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3527..3546
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3681..4475
                     /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(3735..3754)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4345..4364
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4649..4770
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4686..4705)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4740..4759
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4819..4835)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4819..4835)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4832..4854)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4843..4859
                     /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(4867..4873,4874..4891,4892..4897))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4912..4933
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5050..5067)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5221..5809)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5301..5320)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5980..6771,6772..6840))
                     /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     6603..6622
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6841..6945)
                     /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 cggagtcggc
      961 cggagcctcc tccttcttcc cccttgttgt cctcctgctc gccggcagcg gcgggtccgg
     1021 gccccggggg gtccaggctc tgctgtgtgc gtgcaccagc tgcctccagg ccaactacac
     1081 gtgtgagaca gatggggcct gcatggtttc cattttcaat ctggatggga tggagcacca
     1141 tgtgcgcacc tgcatcccca aagtggagct ggtccctgcc gggaagccct tctactgcct
     1201 gagctcggag gacctgcgca acacccactg ctgctacact gactactgca acaggatcga
     1261 cttgagggtg cccagtggtc acctcaagga gcctgagcac ccgtccatgt ggggcccggt
     1321 ggagctggta ggcatcatcg ccggcccggt gttcctcctg ttcctcatca tcatcattgt
     1381 tttccttgtc attaactatc atcagcgtgt ctatcacaac cgccagagac tggacatgga
     1441 agatccctca tgtgagatgt gtctctccaa agacaagacg ctccaggatc ttgtctacga
     1501 tctctccacc tcagggtctg gctcagggtt acccctcttt gtccagcgca cagtggcccg
     1561 agacatcgtt ttacaagaga ttattggcaa gggtcggttt ggggaagtat ggcggggccg
     1621 ctggaggggt ggtgatgtgg ctgtgaaaat attctcttct cgtgaagaac ggtcttggtt
     1681 cagggaagca gagatatacc agacggtcat gctgcgccat gaaaacatcc ttggatttat
     1741 tgctgctgac aataaagata atggcacctg gacacagctg tggcttgttt ctgactatca
     1801 tgagcacggg tccctgtttg attatctgaa ccggtacaca gtgacaattg aggggatgat
     1861 taagctggcc ttgtctgctg ctagtgggct ggcacacctg cacatggaga tcgtgggcac
     1921 ccaagggaag cctggaattg ctcatcgaga cttaaagtca aagaacattc tggtgaagaa
     1981 aaatggcatg tgtgccatag cagacctggg cctggctgtc cgtcatgatg cagtcactga
     2041 caccattgac attgccccga atcagagggt ggggaccaaa cgatacatgg cccctgaagt
     2101 acttgatgaa accattaata tgaaacactt tgactccttt aaatgtgctg atatttatgc
     2161 cctcgggctt gtatattggg agattgctcg aagatgcaat tctggaggag tccatgaaga
     2221 atatcagctg ccatattacg acttagtgcc ctctgaccct tccattgagg aaatgcgaaa
     2281 ggttgtatgt gatcagaagc tgcgtcccaa catccccaac tggtggcaga gttatgaggc
     2341 actgcgggtg atggggaaga tgatgcgaga gtgttggtat gccaacggcg cagcccgcct
     2401 gacggccctg cgcatcaaga agaccctctc ccagctcagc gtgcaggaag acgtgaagat
     2461 cctcgagggt tatccgtatg atgttcctga ttatgctagc ctctaggtcg actctagagg
     2521 gccctattct atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt
     2581 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     2641 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     2701 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     2761 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     2821 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     2881 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     2941 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca
     3001 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3061 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3121 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     3181 cggtctattc ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg
     3241 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     3301 tggaaagtcc ccaggctccc caggcaggca gaagtatgca aagcatgcat ctcaattagt
     3361 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3421 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3481 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3541 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3601 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     3661 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3721 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3781 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3841 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     3901 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     3961 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4021 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4081 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4141 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4201 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4261 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4321 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4381 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4441 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4501 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4561 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4621 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     4681 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4741 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     4801 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4861 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4921 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4981 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5041 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5101 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5161 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5221 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5281 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5341 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5401 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5461 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5521 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5581 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5641 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     5701 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5761 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5821 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5881 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5941 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6001 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6061 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6121 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6181 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6241 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6301 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6361 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6421 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6481 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6541 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6601 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6661 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6721 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6781 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6841 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6901 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6961 agtgccacct gacgtc
//
