LOCUS       sequence-340563-        7919 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses BMPR-2 resistant to the gRNAs(#163388-90) in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7919)
  AUTHORS   Aihara S, Fujimoto S, Sakaguchi R, Imai T
  TITLE     BMPR-2 gates activity-dependent stabilization of primary dendrites 
            during mitral cell remodeling.
  JOURNAL   Cell Rep. 2021 Jun 22;35(12):109276. doi: 
            10.1016/j.celrep.2021.109276.
  PUBMED    34161760
REFERENCE   2  (bases 1 to 7919)
  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: "Cell Rep.";
            date: "2021-06-22"; volume: "35(12):109276. doi"; pages: " 
            10.1016/j.celrep.2021.109276"
COMMENT     Sequence Label: pCAG-Bmpr2 gRNA resistant
FEATURES             Location/Qualifiers
     source          1..7919
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(4941..4960)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5006..5061
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5060..5079)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5422..5438)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5422..5438)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5435..5457)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5446..5462
                     /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(5470..5476,5477..5494,5495..5500))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5515..5536
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5595..5790
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5641..5776
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5796..5930
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5833..5852)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5887..5906
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5998..6015)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6169..6757)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6249..6268)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6928..7719,7720..7788))
                     /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     7551..7570
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7789..7893)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggsgcgc gccaggsggg gsggggsggg gsgrggggsg
      541 gggsggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcgccaccat
     1741 gacttcaagt ctacacagac cattcagagt gccctggctg ctatgggccg tcctgctggt
     1801 cagcactacg gctgcttctc agaatcaaga acggctgtgt gcatttaaag atccatatca
     1861 acaagatctt gggataggtg agagtcgaat ctctcatgaa aatgggacaa tattatgtag
     1921 taagggatca acatgctacg gtctgtggga gaaatcaaaa ggggacatca atcttgtgaa
     1981 acaaggatgt tggtctcaca tcggtgatcc ccaagagtgc cactatgaag agtgtgtagt
     2041 aactaccacc ccaccctcaa ttcagaatgg aacgtaccgc ttttgctgct gtagtacaga
     2101 tttatgtaat gtcaacttta ctgagaactt tccaccccct gacacaacac cactcagtcc
     2161 acctcattca tttaatcgag atgaaacgat aatcattgct ttggcatcag tttctgtgtt
     2221 agctgttttg atagtcgcct tatgttttgg atacagaatg ttgacaggag accggaaaca
     2281 gggtcttcac agcatgaaca tgatggaggc ggcagcagca gagccctccc ttgacctgga
     2341 taacctgaag ctgctggagc tgattggacg gggtaggtat ggtgctgttt acaagggttc
     2401 cttggatgag cgtccagttg ctgtaaaagt attttctttt gcaaaccgtc agaattttat
     2461 aaatgaaaaa aacatttaca gagtgccttt gatggaacat gacaacattg ctcgcttcat
     2521 agttggagac gagaggctca ctgcagacgg ccgcatggag tatttgcttg tgatggagta
     2581 ttatcccaat ggatctctgt gcaaatatct gagtctccac acaagtgatt gggtaagctc
     2641 ttgccgtctg gctcattctg tgactagagg actggcttat cttcacacag aattaccacg
     2701 aggagatcat tataaacccg caatctccca ccgagattta aacagcagga atgtcctggt
     2761 aaagaatgac ggcgcgtgtg ttatcagtga ctttggttta tccatgaggc taactggaaa
     2821 tcggctggtg cgcccagggg aagaagataa tgcggctata agtgaggttg gcacaattcg
     2881 ctatatggca ccagaagtgc tagaaggagc tgtgaacctg agggactgtg agtcagctct
     2941 gaagcaagtg gacatgtatg cgcttggact catctactgg gaggtgttta tgaggtgtac
     3001 agacctcttc ccaggtgaat ctgtaccaga ttaccagatg gcttttcaga cagaagttgg
     3061 aaaccatccc acatttgagg atatgcaggt tcttgtgtcc agagagaagc agagacccaa
     3121 gttcccagaa gcctggaaag aaaatagcct ggcagtgagg tcactcaagg aaacaattga
     3181 agactgctgg gaccaggatg cagaggctcg gctcactgca cagtgtgctg aggagaggat
     3241 ggctgaactc atgatgatat gggagagaaa caagtctgtg agcccaacgg tcaacccaat
     3301 gtcaactgct atgcagaatg aacgcaacct gtcacataat aggcgtgtgc caaaaatcgg
     3361 gccttaccca gattattcct cttcctcata tattgaagac tctatccatc atactgacag
     3421 cattgtgaag aatatttcct ctgagcattc gatgtccagc acaccattga caataggaga
     3481 aaagaatcga aattcaatta attatgaacg acagcaagca caagctcgaa tccctagccc
     3541 agaaacaagc gtcacaagcc tgtccacaaa cacaaccacc acaaacacca ccggcctcac
     3601 tccaagtact ggcatgacca ctatatctga gatgccatac ccagatgaga cacatttgca
     3661 cgccacaaat gttgcacagt caatcgggcc aacccctgtc tgcttacagc tgacagaaga
     3721 agacttggag actaataagc tagatccaaa agaagttgat aagaacctca aggaaagctc
     3781 tgatgagaat ctcatggagc attctctgaa gcagttcagt gggccagacc cattgagcag
     3841 taccagttct agcttgcttt atccactcat aaagctcgca gtggaagtga ctggacaaca
     3901 ggacttcaca caggctgcaa atgggcaagc atgtttaatt cctgatgttc cacctgctca
     3961 gatctatcct ctccctaagc aacagaacct tcctaagaga cctactagtt tgcctttgaa
     4021 caccaaaaat tcaacaaaag aaccccggct aaaatttggc aacaagcaca aatcaaactt
     4081 gaaacaagta gaaactggag ttgccaagat gaatacaatc aatgcagcag agcctcatgt
     4141 ggtgacagta actatgaatg gtgtggcagg tagaagccac aatgttaatt cccatgctgc
     4201 cacaacccag tatgccaatg gcgcagtgcc agctggccag gcagccaaca tagtggcaca
     4261 taggtcccaa gaaatgctgc agaatcaatt tattggtgag gataccaggc tgaatatcaa
     4321 ttccagtcct gatgagcatg aacctttact gagacgagag caacaggctg gccatgatga
     4381 aggggttctg gatcgtttgg tagataggag ggaacggcca ttagaaggtg gccgaacaaa
     4441 ttccaataac aacaacagca atccatgttc agaacaagat atccttacac aaggtgttac
     4501 aagcacagct gcagatcctg ggccatcaaa gcccagaaga gcacagaggc ccaattctct
     4561 ggatctttca gccacaaata tcctggatgg cagcagtata cagataggtg agtcaacaca
     4621 agatggcaaa tcaggatcag gtgaaaagat caagagacgt gtgaaaactc catactctct
     4681 taagcggtgg cgcccgtcca cctgggtcat ctccaccgag ccgctggact gtgaggtcaa
     4741 caacaatggc agtgacaggg cagtccattc taaatctagc actgctgtgt accttgcaga
     4801 gggaggcact gccacgacca cagtgtctaa agatatagga atgaattgtc tgtgagcggc
     4861 cgcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg
     4921 gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag
     4981 ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt
     5041 tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat
     5101 cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac
     5161 aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta
     5221 ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt
     5281 ttttctttaa catccctaaa attttcctta catgttttac tagccagatt tttcctcctc
     5341 tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcag
     5401 cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5461 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5521 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5581 gtgccagcgg atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     5641 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     5701 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     5761 ggcttttttg gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg
     5821 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5881 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta
     5941 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     6001 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     6061 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     6121 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6181 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6241 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6301 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6361 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6421 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6481 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6541 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6601 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6661 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6721 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6781 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6841 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6901 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6961 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     7021 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7081 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7141 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7201 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     7261 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7321 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7381 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7441 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7501 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7561 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7621 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7681 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7741 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7801 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7861 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctg
//
