LOCUS       sequence-456512-        6899 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of ALK5 WT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6899)
  AUTHORS   Moren A, Imamura T, Miyazono K, Heldin CH, Moustakas A
  TITLE     Degradation of the tumor suppressor Smad4 by WW and HECT domain 
            ubiquitin ligases.
  JOURNAL   J Biol Chem. 2005 Jun 10;280(23):22115-23. Epub 2005 Apr 6.
  PUBMED    15817471
REFERENCE   2  (bases 1 to 6899)
  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: "J Biol 
            Chem."; date: "2005-06-10"; volume: "280(23)"; pages: "22115-23. 
            Epub 2005 Apr 6"
COMMENT     Sequence Label: pcDNA3-ALK5 wt
FEATURES             Location/Qualifiers
     source          1..6899
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(488..507)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(489..507)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(552..755)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(581..601)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        756..1135
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1307..1326
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1401..1505
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1506..1574,1575..2366)
                     /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     complement(1724..1743)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2537..3125
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3026..3045
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3279..3296
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3413..3434
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3449..3454,3455..3472,3473..3479)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3487..3503
                     /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)."
     primer_bind     3492..3514
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3511..3527
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3511..3527
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3576..3697
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3587..3606)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3641..3660
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3871..4665)
                     /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(3982..4001)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4592..4611
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4732..4928)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        complement(4740..4950)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4746..4881
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4800..4819)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     4974..4994
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(5003..5431)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5113..5134)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5325..5344
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    5477..5701
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5690..5707
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        5727..5745
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5727..5744
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(5770..5796)
                     /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"
     CDS             complement(5776..5802)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
ORIGIN
        1 gtgagatgca gacgaagcac actggtccag caatgacagc tgccagttcc acaggaccaa
       61 ggccaggtga tgactttaca gtagttggaa gttctatttt attgcaatgg tcctgattgc
      121 agcaatatgt tgtagtcaca gacccagttt ttgaagaggg tgcacataca aacggcctat
      181 ctcgtggaat taagtcaatt tcagctatac acatgctgtt gtgtataact ttgtctgtgg
      241 tctctgtgac agagacaaag cagagcccat ctgtcacaca agtaaaattg tcttttgtac
      301 agaggtggca gaaacactgt aacgccgtcg cccccgggag cagcgccgcc gccgccgccg
      361 ccgccgccgc cagcacgagg aggagcagcc ggggacgcgg agcagcgacc gccgcctcca
      421 tggcccgaat tccagcacac tggcggccgt tactagtgga tccgagctcg gtaccaagct
      481 tgggtctccc tatagtgagt cgtattaatt tcgataagcc agtaagcagt gggttctcta
      541 gttagccaga gagctctgct tatatagacc tcccaccgta cacgcctacc gcccatttgc
      601 gtcaatgggg cggagttgtt acgacatttt ggaaagtccc gttgattttg gtgccaaaac
      661 aaactcccat tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc aaaccgctat
      721 ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg taatagcgat gactaatacg
      781 tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg
      841 ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt
      901 actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct
      961 attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc
     1021 ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta
     1081 tgaactaatg accccgtaat tgattactat taataactag tcaataatca atgtcaacgc
     1141 gtatatctgg cccgtacatc gcgaagcagc gcaaaacgcc taaccctaag cagattcttc
     1201 atgcaattgt cggtcaagcc ttgccttgtt gtagcttaaa ttttgctcgc gcactactca
     1261 gcgacctcca acacacaagc agggagcaga tactggctta actatgcggc atcagagcag
     1321 attgtactga gagtgcacca taggggatcg ggagatctcc cgatccgtcg acgtcaggtg
     1381 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     1441 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     1501 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     1561 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     1621 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     1681 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     1741 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     1801 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     1861 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     1921 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     1981 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     2041 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     2101 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     2161 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     2221 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     2281 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     2341 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     2401 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     2461 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     2521 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     2581 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     2641 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     2701 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     2761 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     2821 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     2881 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     2941 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3001 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3061 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     3121 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     3181 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     3241 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     3301 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     3361 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     3421 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     3481 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     3541 agctctagct agaggtcgac ggtatacaga catgataaga tacattgatg agtttggaca
     3601 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     3661 tttatttgta accattataa gctgcaataa acaagttggg gtgggcgaag aactccagca
     3721 tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca
     3781 acctttcata gaaggcggcg gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt
     3841 ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa
     3901 ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca
     3961 ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc
     4021 cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat
     4081 attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc
     4141 cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc
     4201 ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg
     4261 gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat
     4321 gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc
     4381 gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg
     4441 aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcctgcagtt cattcagggc
     4501 accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac
     4561 ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac
     4621 ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca
     4681 tcctgtctct tgatcagatc cgaaaatgga tatacaagct cccgggagct ttttgcaaaa
     4741 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg cagaggcggc
     4801 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
     4861 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
     4921 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa
     4981 ctgacacaca ttccacagaa ttaattcgcg ttaaattttt gttaaatcag ctcatttttt
     5041 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     5101 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     5161 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     5221 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     5281 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     5341 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     5401 gccgcgctta atgcgccgct acagggcgcg tggggatacc ccctagagcc ccagctggtt
     5461 ctttccgcct cagaagccat agagcccacc gcatccccag catgcctgct attgtcttcc
     5521 caatcctccc ccttgctgtc ctgccccacc ccacccccca gaatagaatg acacctactc
     5581 agacaatgcg atgcaatttc ctcattttat taggaaagga cagtgggagt ggcaccttcc
     5641 agggtcaagg aaggcacggg ggaggggcaa acaacagatg gctggcaact agaaggcaca
     5701 gtcgaggctg atcagcgagc tctagcattt aggtgacact atagaatagg gccctctaga
     5761 gtcgacctag aggctagcat aatcaggaac atcatacgga taaccctcga gcattttgat
     5821 gccttcctgt tgactgagtt gcgataatgt tttcttaatc cgcaatgctg taagcctagc
     5881 tgctccattg gcataccaac attctctcat aattttagcc attactctca aggcttcaca
     5941 gctctgccat ctgtttggga tatttggcct taacttctgt tcacaaacaa cttttctcat
     6001 ttcttcaact gatgggtcag aaggtacaag atcataataa ggcagttggt aatcttcatg
     6061 aataccacca atggaacatc gtcgagcaat ttcccagaat actaagccca ttgcatagat
     6121 gtcagcacgt ttgaaggatt caaaatgttt catatttatg gaatcatcga gaacttcagg
     6181 ggccatgtac ctttttgttc ccactctgtg gtttggagca atatcaatgg tatctgtggc
     6241 tgaatcatgt cttactgcca gtcctaagtc tgcaatacag caagttccat tcttctttac
     6301 caagatattc tttgatttca aatctctatg agcaatggct ggctttcctt gggtaccaac
     6361 aatctccatg tgaagatggg caagaccgct cgccgtggac agagcaagtt ttatcattcc
     6421 ttccacagta actgtgtatc tgtttaagta atcaaaaagg gatccatgct catgataatc
     6481 tgacaccaac cagagctgag tccaagtacc attgtcttta ttgtctgctg ctataaatcc
     6541 caggatgttt tcatgacgta acattacagt ttgataaatc tctgcctcac ggaaccacga
     6601 acgttcttct ctagaggaga atatcttaac agcaacttct tctccccgcc actttcctct
     6661 ccaaacttct ccaaatcgac ctttgccaat gctttcttgt aacacaatag ttctcgcaat
     6721 tgttctctga acaagcaatg gtaaacctga gccagaacct gacgttgtca tatcataaat
     6781 taagtctttc aacgtagtac cctctgaaat aaaagggcga tctaatgaag ggtcctcttc
     6841 atttggcact cgatggtgaa tgacagtgcg gttgtggcag atatagacca tcaacatga
//
