LOCUS       sequence-395195-        5678 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  in vitro transcription of RNA encoding HA-tagged Acvr1l that 
            contains 4 amino acid substitutions: T186V, S187A, S189A, S191A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5678)
  AUTHORS   Tajer B, Dutko JA, Little SC, Mullins MC
  TITLE     BMP heterodimers signal via distinct type I receptor class 
            functions.
  JOURNAL   Proc Natl Acad Sci U S A. 2021 Apr 13;118(15):e2017952118. doi: 
            10.1073/pnas.2017952118.
  PUBMED    33827919
REFERENCE   2  (bases 1 to 5678)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2021-04-13"; volume: "118(15):e2017952118. 
            doi"; pages: " 10.1073/pnas.2017952118"
COMMENT     Sequence Label: acvr1l-VAAA-HA
FEATURES             Location/Qualifiers
     source          1..5678
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1650..1676
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1730..1864
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1754..1773)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1808..1827
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1977..1997)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1977..1995)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2016..2032)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2016..2032)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2029..2051)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2040..2056
                     /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(2064..2070,2071..2088,2089..2094))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2109..2130
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2247..2264)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2418..3006)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2498..2517)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3177..3968,3969..4037))
                     /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     3800..3819
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4038..4142)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4168..4623)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4305..4326)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4517..4536
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4694..5678
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccggcagc gctgagagtc acttgaggag ttgtacaatg
      121 cattgaacca aatggggcat tgcagcaccc aaatcatcat cctgttcctg cttcagttct
      181 tacagacatc agctaaagat gtctccattg actgcatgtg tgtcggcagt gactgtaatg
      241 agcagcagtg tactggtgac cagtgttaca cctccgttat cattagcaat gatgtgacga
      301 cgttcaagcg gggctgcttg atcgggccgg cgagcaagcg catgacctgc tccgcaacag
      361 cttctgctag tcatgtggta gaatgctgtt ctcaacacat gtgcaacgcc aacgtctcca
      421 aagagaccct acttcgactg ctgctcacaa gtccagaaga aaagaagact gttcattacc
      481 gcgtggaaat gttggttctg tttgtgttgg ggccgtttgt ggttctgggt ctgctgtctt
      541 ttctggcttt gctggtgtgc cgtcgactcc atcatgggcg tctggagaga ctgcacgagt
      601 ttgacactga acagggggcc atcgatgggc ttatcgcgtc taatgtcgga gacagcacac
      661 ttgcggatct gatggatcac tcctgcgttg caggcgctgg tgcaggactg cccttcctgg
      721 ttcagagaac ggttgcgcgg cagatcagcc tggtggagtg tgttggtaaa ggacggtacg
      781 gtgaagtgtg gagaggtcaa tggcaaggag aaaatgtagc cgtgaagatc ttttcctcta
      841 gagatgagaa gtcatggttt cgagaaacag aaatttacaa cactgttctg ctacgacatg
      901 aaaatatatt aggcttcatg gcttctgaca tgacctcccg aaactctagc actcagctgt
      961 ggctgatcac acactatcac gagaatggct ctctgtatga ctacctgcag cgtgtggctg
     1021 tggagatggc agatggactg cacatggcgg cttcgattgc cagcgggctg gtgcacctgc
     1081 acacggagat ctttggcacg gagggcaaac cggccatcgc tcacagagac ctgaagagca
     1141 agaacatcct tgtgaagaaa gatttgcagt gctgcatcgc tgacctgggt ctggcagtaa
     1201 cacacacgca gtctgataat cagcttgatg tgggaaataa tcctaaagtg ggaaccaaac
     1261 gctacatggc accggaggtt ctagatgaga ccattcagac ggactgtttt gacgcctata
     1321 agagggtgga tatctgggcc tttgggttgg tgctgtggga gatcgcacgc agaaccatca
     1381 gcaatggaat tgtagaggaa tacaagccgc ctttctatga cctggttcct aatgatccca
     1441 gctttgacga catgaggaaa gtggtttgtg tggagcagca aaggccattc attcccaacc
     1501 gctggttttc agatcctacc ctgtctgctc tggtgaagct gatgaaagag tgctggtacc
     1561 agaacccctc ggctcgtctc actgccctgc gcatcaaaaa gactctggat aaaatccaca
     1621 gttcactgga gaagggcaaa accgactgct acccatacga tgtgccagat tacgcttgag
     1681 aattcaaggc ctctcgagcc tctagaacta tagtgagtcg tattacgtag atccagacat
     1741 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     1801 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     1861 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     1921 tttttaattc gcggccgcgg cgccaatgca ttgggcccgg tacccagctt ttgttccctt
     1981 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     2041 tgttatccgc tcacaattcc acacaacata cgagccggga gcataaagtg taaagcctgg
     2101 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     2161 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     2221 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     2281 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2341 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2401 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2461 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2521 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2581 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2641 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2701 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2761 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2821 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2881 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2941 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3001 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3061 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3121 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3181 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3241 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3301 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3361 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3421 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     3481 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     3541 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     3601 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3661 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3721 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3781 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     3841 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     3901 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     3961 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     4021 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     4081 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     4141 cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg
     4201 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     4261 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     4321 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     4381 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     4441 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     4501 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     4561 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     4621 cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     4681 cgctattacg ccagtcgacc gccaattcaa tatggcgtat atggactcat gccaattcaa
     4741 tatggtggat ctggacctgt gccaattcaa tatggcgtat atggactcgt gccaattcaa
     4801 tatggtggat ctggacccca gccaattcaa tatggcggac ttggcaccat gccaattcaa
     4861 tatggcggac ctggcactgt gccaactggg gaggggtcta cttggcacgg tgccaagttt
     4921 gaggaggggt cttggccctg tgccaagtcc gccatattga attggcatgg tgccaataat
     4981 ggcggccata ttggctatat gccaggatca atatataggc aatatccaat atggccctat
     5041 gccaatatgg ctattggcca ggttcaatac tatgtattgg ccctatgcca tatagtattc
     5101 catatatggg ttttcctatt gacgtagata gcccctccca atgggcggtc ccatatacca
     5161 tatatggggc ttcctaatac cgcccatagc cactccccca ttgacgtcaa tggtctctat
     5221 atatggtctt tcctattgac gtcatatggg cggtcctatt gacgtatatg gcgcctcccc
     5281 cattgacgtc aattacggta aatggcccgc ctggctcaat gcccattgac gtcaatagga
     5341 ccacccacca ttgacgtcaa tgggatggct cattgcccat tcatatccgt tctcacgccc
     5401 cctattgacg tcaatgacgg taaatggccc acttggcagt acatcaatat ctattaatag
     5461 taacttggca agtacattac tattggaagt acgccagggt acattggcag tactcccatt
     5521 gacgtcaatg gcggtaaatg gcccgcgatg gctgccaagt acatccccat tgacgtcaat
     5581 ggggaggggc aatgacgcaa atgggcgttc cattgacgta aatgggcggt aggcgtgcct
     5641 aatgggaggt ctatataagc aatgctcgtt tagggaac
//
