LOCUS       sequence-395422-        5408 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  in vitro transcription of RNA encoding FLAG-tagged Bmp7a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5408)
  AUTHORS   Little SC, Mullins MC
  TITLE     Bone morphogenetic protein heterodimers assemble heteromeric type I 
            receptor complexes to pattern the dorsoventral axis.
  JOURNAL   Nat Cell Biol. 2009 May;11(5):637-43. doi: 10.1038/ncb1870. Epub 
            2009 Apr 19.
  PUBMED    19377468
REFERENCE   2  (bases 1 to 5408)
  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"; doi: "10.1038/ncb1870"; 
            journalName: "Nat Cell Biol"; date: "2009-05"; volume: "11"; issue: 
            "5"; pages: "637-43"
COMMENT     Sequence Label: FLAG-Bmp7
FEATURES             Location/Qualifiers
     source          1..5408
                     /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             1015..1038
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1460..1594
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1484..1503)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1538..1557
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1707..1727)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1707..1725)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1746..1762)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1746..1762)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1759..1781)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1770..1786
                     /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(1794..1800,1801..1818,1819..1824))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1839..1860
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1977..1994)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2148..2736)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2228..2247)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2907..3698,3699..3767))
                     /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     3530..3549
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3768..3872)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3898..4353)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4035..4056)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4247..4266
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4424..5408
                     /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 atcaatgact ctaaaaatgt ttccagtgca tcttatttcc
      121 actttgatag tggcttgtgg ttgcagtgtg ctcgcagacg caatgcaagc aaacttcacc
      181 atggataatg acattcagtc cagcttcatc cagcggcgcc tgaagagcca ggagcgcaga
      241 gagatgcagc gggaaatcct ctctatcctc gggctgccgc aacggccacg tccgctcctg
      301 cacgagagac acaccgcggc tcccatgtac atgctggatc tgtataacgc catcctggag
      361 gacggagaca ggcgcggtgg gcttgtgtac tcttatgaac ccgcgtacac gaccccggga
      421 cccccgctgg tgacccagca ggacagtcgc tttctcagtg atgccgacat ggtgatgagc
      481 tttgcgaata cagtggatcc agaagaagac cttcaactgt atcatcagca tcgcagagag
      541 ttcagatttg acctttctcg catcccgcca ggtgaaactg tgaccgctgc agagttccga
      601 atctacaaag actttgttcg cgagcgttat gaaaacgaga ccttccatgt cagcgtgttc
      661 caggtgcttc aacagcagca cagaagggag ctgtacctgc tggactcacg agtggtatgg
      721 gctgcagaag agggatggct ggtctttgac ctcacggtca ccagtaacca ctgggttata
      781 aacccaggtc agaacttagg actgcagctc ttagtggaga cctcacatgg tgcaagaatg
      841 aacccaagaa gagctggcct ggtgggcagc agtggagctc agaataagca gccgtttatg
      901 gtggcgtttt tgaaagcatc aggaatccac ctccgcagtg tccgctcggc atcaggaggc
      961 aaacagaagg gtcatcaccg cacaaaaaac gccaaacctg gcgctggagg cggagattac
     1021 aaggatgatg atgataaggg aggcggagct gcacacagcc aggtggcttt aaaaacagct
     1081 gaagccacag agggtgccag catagatccc aaacagggct gcaaaaagca tgaactctac
     1141 gtcagcttca gagatctggg atggcaggac tggatcatcg ccccagaggg ttatgccgcg
     1201 tactactgtg agggcgaatg tgtgttccct ttgaactctt atatgaacgc cacaaaccat
     1261 gctattgtac agacactggt tcatttcata aaccctgaaa ctgtcccgaa gccctgctgc
     1321 gctccaactc agctgcacgg aatctcagtt ttgtacttcg atgacagctc caacgtaata
     1381 ttgaaaaagt accgcaacat ggtcgtcaga gcctgcggat gccactaatc tctagaacta
     1441 tagtgagtcg tattacgtag atccagacat gataagatac attgatgagt ttggacaaac
     1501 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     1561 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     1621 gtttcaggtt cagggggagg tgtgggaggt tttttaattc gcggccgcgg cgccaatgca
     1681 ttgggcccgg tacccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
     1741 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     1801 cgagccggga gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     1861 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     1921 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     1981 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2041 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2101 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2161 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2221 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2281 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2341 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2401 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2461 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2521 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2581 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2641 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2701 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2761 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2821 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     2881 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     2941 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3001 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3061 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3121 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3181 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3241 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3301 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3361 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3421 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     3481 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     3541 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     3601 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     3661 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     3721 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     3781 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     3841 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa
     3901 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     3961 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     4021 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     4081 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     4141 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     4201 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     4261 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     4321 ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact
     4381 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagtcgacc gccaattcaa
     4441 tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt gccaattcaa
     4501 tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca gccaattcaa
     4561 tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt gccaactggg
     4621 gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg tgccaagtcc
     4681 gccatattga attggcatgg tgccaataat ggcggccata ttggctatat gccaggatca
     4741 atatataggc aatatccaat atggccctat gccaatatgg ctattggcca ggttcaatac
     4801 tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt gacgtagata
     4861 gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac cgcccatagc
     4921 cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac gtcatatggg
     4981 cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta aatggcccgc
     5041 ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa tgggatggct
     5101 cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg taaatggccc
     5161 acttggcagt acatcaatat ctattaatag taacttggca agtacattac tattggaagt
     5221 acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg gcccgcgatg
     5281 gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa atgggcgttc
     5341 cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc aatgctcgtt
     5401 tagggaac
//
