LOCUS       sequence-406401-        5674 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  in vitro transcription of RNA encoding human ACVR1 with mutations 
            S190A, S192A, and R206H.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5674)
  AUTHORS   Allen RS, Jones WD, Hale M, Warder BN, Shore EM, Mullins MC
  TITLE     Reduced GS domain serine/threonine requirements of Fibrodysplasia 
            Ossificans Progressiva mutant type I BMP receptor ACVR1 in the 
            zebrafish.
  JOURNAL   J Bone Miner Res. 2023 Jun 17. doi: 10.1002/jbmr.4869.
  PUBMED    37329499
REFERENCE   2  (bases 1 to 5674)
  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 Bone 
            Miner Res. 2023 Jun 17. doi: 10.1002/jbmr.4869."
COMMENT     Sequence Label: ACVR1-TAAS-R206H
FEATURES             Location/Qualifiers
     source          1..5674
                     /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             1630..1677
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     polyA_signal    1727..1861
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1751..1770)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1805..1824
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1974..1994)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1974..1992)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2013..2029)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2013..2029)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2026..2048)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2037..2053
                     /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(2061..2067,2068..2085,2086..2091))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2106..2127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2244..2261)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2414..3002)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2494..2513)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3173..3964,3965..4033))
                     /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     3796..3815
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4034..4138)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4164..4619)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4301..4322)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4513..4532
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4690..5674
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctgggaacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccatggta gatggagtga tgattcttcc tgtgcttatc
      121 atgattgctc tcccctcccc tagtatggaa gatgagaagc ccaaggtcaa tcccaaactc
      181 tacatgtgtg tgtgtgaagg tctctcctgc ggtaatgagg accactgtga aggccagcag
      241 tgcttttcct cactgagcat caacgatggc ttccacgtct accagaaagg ctgcttccag
      301 gtttatgagc agggaaagat gacctgtaag accccgccgt cccctggcca agctgtggag
      361 tgctgccaag gggactggtg taacaggaac atcacggccc agctgcccac taaaggaaaa
      421 tccttccctg gaacacagaa tttccacttg gaggttggcc tcattattct ctctgtagtg
      481 ttcgcagtat gtcttttagc ctgcctgctg ggagttgctc tccgaaaatt taaaaggcgc
      541 aaccaagaac gcctcaatcc ccgagacgtg gagtatggca ctatcgaagg gctcatcacc
      601 accaatgttg gagacagcac tttagcagat ttattggatc attcgtgtac agcaggagct
      661 ggctctggtc ttccttttct ggtacaaaga acagtggctc accagattac actgttggag
      721 tgtgtcggga aaggcaggta tggtgaggtg tggaggggca gctggcaagg ggaaaatgtt
      781 gccgtgaaga tcttctcctc ccgtgatgag aagtcatggt tcagggaaac ggaattgtac
      841 aacactgtga tgctgaggca tgaaaatatc ttaggtttca ttgcttcaga catgacatca
      901 agacactcca gtacccagct gtggttaatt acacattatc atgaaatggg atcgttgtac
      961 gactatcttc agcttactac tctggataca gttagctgcc ttcgaatagt gctgtccata
     1021 gctagtggtc ttgcacattt gcacatagag atatttggga cccaagggaa accagccatt
     1081 gcccatcgag atttaaagag caaaaatatt ctggttaaga agaatggaca gtgttgcata
     1141 gcagatttgg gcctggcagt catgcattcc cagagcacca atcagcttga tgtggggaac
     1201 aatccccgtg tgggcaccaa gcgctacatg gcccccgaag ttctagatga aaccatccag
     1261 gtggattgtt tcgattctta taaaagggtc gatatttggg cctttggact tgttttgtgg
     1321 gaagtggcca ggcggatggt gagcaatggt atagtggagg attacaagcc accgttctac
     1381 gatgtggttc ccaatgaccc aagttttgaa gatatgagga aggtagtctg tgtggatcaa
     1441 caaaggccaa acatacccaa cagatggttc tcagacccga cattaacctc tctggccaag
     1501 ctaatgaaag aatgctggta tcaaaatcca tccgcaagac tcacagcact gcgtatcaaa
     1561 aagactttga ccaaaattga taattccctc gacaaattga aaactgactg taatcgattt
     1621 aaagctatgg attacaagga tgatgatgat aaggattaca aggatgatga tgataagaat
     1681 tcaaggcctc tcgagcctct agaactatag tgagtcgtat tacgtagatc cagacatgat
     1741 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     1801 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     1861 taacaacaac aattgcattc attttatgtt acaggttcag ggggaggtgt gggaggtttt
     1921 ttaattcgcg gccgcggcgc caatgcattg ggcccggtac ccagcttttg ttccctttag
     1981 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     2041 tatccgctca caattccaca caacatacga gccgggagca taaagtgtaa agcctggggt
     2101 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     2161 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     2221 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2281 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2341 aacgcaggaa agaacatgtg agcaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2401 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2461 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2521 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2581 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2641 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2701 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2761 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2821 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2881 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2941 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3001 aagaagaccc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3061 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3121 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3181 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3241 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3301 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3361 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3421 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3481 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3541 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     3601 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     3661 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     3721 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     3781 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     3841 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3901 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     3961 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4021 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4081 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4141 catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt
     4201 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     4261 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     4321 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     4381 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     4441 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     4501 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     4561 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     4621 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     4681 attacgccag tcgaccgcca attcaatatg gcgtatatgg actcatgcca attcaatatg
     4741 gtggatctgg acctgtgcca attcaatatg gcgtatatgg actcgtgcca attcaatatg
     4801 gtggatctgg accccagcca attcaatatg gcggacttgg caccatgcca attcaatatg
     4861 gcggacctgg cactgtgcca actggggagg ggtctacttg gcacggtgcc aagtttgagg
     4921 aggggtcttg gccctgtgcc aagtccgcca tattgaattg gcatggtgcc aataatggcg
     4981 gccatattgg ctatatgcca ggatcaatat ataggcaata tccaatatgg ccctatgcca
     5041 atatggctat tggccaggtt caatactatg tattggccct atgccatata gtattccata
     5101 tatgggtttt cctattgacg tagatagccc ctcccaatgg gcggtcccat ataccatata
     5161 tggggcttcc taataccgcc catagccact cccccattga cgtcaatggt ctctatatat
     5221 ggtctttcct attgacgtca tatgggcggt cctattgacg tatatggcgc ctcccccatt
     5281 gacgtcaatc acggtaaatg gcccgcctgg ctcaatgccc attgacgtca ataggaccac
     5341 ccaccattga cgtcaatggg atggctcatt gcccattcat atccgttctc acgcccccta
     5401 ttgacgtcaa tgacggtaaa tggcccactt ggcagtacat caatatctat taatagtaac
     5461 ttggcaagta cattactatt ggaagtacgc cagggtacat tggcagtact cccattgacg
     5521 tcaatggcgg taaatggccc gcgatggctg ccaagtacat ccccattgac gtcaatgggg
     5581 aggggcaatg acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc gtgcctaatg
     5641 ggaggtctat ataagcaatg ctcgtttagg gaac
//
