LOCUS       sequence-339003-        7548 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses BMPR-2-Delta-EC 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 7548)
  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 7548)
  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-Delta-EC gRNA resistant
FEATURES             Location/Qualifiers
     source          1..7548
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1687..1706
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(4570..4589)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4635..4690
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4689..4708)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5051..5067)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5051..5067)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5064..5086)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5075..5091
                     /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(5099..5105,5106..5123,5124..5129))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5144..5165
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5224..5419
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5270..5405
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5425..5559
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5462..5481)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5516..5535
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5627..5644)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5798..6386)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5878..5897)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6557..7348,7349..7417))
                     /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     7180..7199
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7418..7522)
                     /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 gggggnsgcg cgccrggsgg ggsggggsgg ggsgrggggs
      541 ggggsggggs gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcgccacca
     1741 tgacttcaag tctacacaga ccattcagag tgccctggct gctatgggcc gtcctgctgg
     1801 tcagcactac ggctgctata atcattgctt tggcatcagt ttctgtgtta gctgttttga
     1861 tagtcgcctt atgttttgga tacagaatgt tgacaggaga ccggaaacag ggtcttcaca
     1921 gcatgaacat gatggaggcg gcagcagcag agccctccct tgacctggat aacctgaagc
     1981 tgctggagct gattggacgg ggtaggtatg gtgctgttta caagggttcc ttggatgagc
     2041 gtccagttgc tgtaaaagta ttttcttttg caaaccgtca gaattttata aatgaaaaaa
     2101 acatttacag agtgcctttg atggaacatg acaacattgc tcgcttcata gttggagacg
     2161 agaggctcac tgcagacggc cgcatggagt atttgcttgt gatggagtat tatcccaatg
     2221 gatctctgtg caaatatctg agtctccaca caagtgattg ggtaagctct tgccgtctgg
     2281 ctcattctgt gactagagga ctggcttatc ttcacacaga attaccacga ggagatcatt
     2341 ataaacccgc aatctcccac cgagatttaa acagcaggaa tgtcctggta aagaatgacg
     2401 gcgcgtgtgt tatcagtgac tttggtttat ccatgaggct aactggaaat cggctggtgc
     2461 gcccagggga agaagataat gcggctataa gtgaggttgg cacaattcgc tatatggcac
     2521 cagaagtgct agaaggagct gtgaacctga gggactgtga gtcagctctg aagcaagtgg
     2581 acatgtatgc gcttggactc atctactggg aggtgtttat gaggtgtaca gacctcttcc
     2641 caggtgaatc tgtaccagat taccagatgg cttttcagac agaagttgga aaccatccca
     2701 catttgagga tatgcaggtt cttgtgtcca gagagaagca gagacccaag ttcccagaag
     2761 cctggaaaga aaatagcctg gcagtgaggt cactcaagga aacaattgaa gactgctggg
     2821 accaggatgc agaggctcgg ctcactgcac agtgtgctga ggagaggatg gctgaactca
     2881 tgatgatatg ggagagaaac aagtctgtga gcccaacggt caacccaatg tcaactgcta
     2941 tgcagaatga acgcaacctg tcacataata ggcgtgtgcc aaaaatcggg ccttacccag
     3001 attattcctc ttcctcatat attgaagact ctatccatca tactgacagc attgtgaaga
     3061 atatttcctc tgagcattcg atgtccagca caccattgac aataggagaa aagaatcgaa
     3121 attcaattaa ttatgaacga cagcaagcac aagctcgaat ccctagccca gaaacaagcg
     3181 tcacaagcct gtccacaaac acaaccacca caaacaccac cggcctcact ccaagtactg
     3241 gcatgaccac tatatctgag atgccatacc cagatgagac acatttgcac gccacaaatg
     3301 ttgcacagtc aatcgggcca acccctgtct gcttacagct gacagaagaa gacttggaga
     3361 ctaataagct agatccaaaa gaagttgata agaacctcaa ggaaagctct gatgagaatc
     3421 tcatggagca ttctctgaag cagttcagtg ggccagaccc attgagcagt accagttcta
     3481 gcttgcttta tccactcata aagctcgcag tggaagtgac tggacaacag gacttcacac
     3541 aggctgcaaa tgggcaagca tgtttaattc ctgatgttcc acctgctcag atctatcctc
     3601 tccctaagca acagaacctt cctaagagac ctactagttt gcctttgaac accaaaaatt
     3661 caacaaaaga accccggcta aaatttggca acaagcacaa atcaaacttg aaacaagtag
     3721 aaactggagt tgccaagatg aatacaatca atgcagcaga gcctcatgtg gtgacagtaa
     3781 ctatgaatgg tgtggcaggt agaagccaca atgttaattc ccatgctgcc acaacccagt
     3841 atgccaatgg cgcagtgcca gctggccagg cagccaacat agtggcacat aggtcccaag
     3901 aaatgctgca gaatcaattt attggtgagg ataccaggct gaatatcaat tccagtcctg
     3961 atgagcatga acctttactg agacgagagc aacaggctgg ccatgatgaa ggggttctgg
     4021 atcgtttggt agataggagg gaacggccat tagaaggtgg ccgaacaaat tccaataaca
     4081 acaacagcaa tccatgttca gaacaagata tccttacaca aggtgttaca agcacagctg
     4141 cagatcctgg gccatcaaag cccagaagag cacagaggcc caattctctg gatctttcag
     4201 ccacaaatat cctggatggc agcagtatac agataggtga gtcaacacaa gatggcaaat
     4261 caggatcagg tgaaaagatc aagagacgtg tgaaaactcc atactctctt aagcggtggc
     4321 gcccgtccac ctgggtcatc tccaccgagc cgctggactg tgaggtcaac aacaatggca
     4381 gtgacagggc agtccattct aaatctagca ctgctgtgta ccttgcagag ggaggcactg
     4441 ccacgaccac agtgtctaaa gatataggaa tgaattgtct gtgagcggcc gcactcctca
     4501 ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg ctcacaaata
     4561 ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca
     4621 tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     4681 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta
     4741 tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct
     4801 ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa
     4861 agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac
     4921 atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct cctgactact
     4981 cccagtcata gctgtccctc ttctcttatg gagatccctc gacctgcagc ccaagcttgg
     5041 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     5101 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     5161 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga
     5221 tccgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     5281 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     5341 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     5401 aggcctaggc ttttgcaaaa agctaacttg tttattgcag cttataatgg ttacaaataa
     5461 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5521 ttgtccaaac tcatcaatgt atcttatcat gtctggatcc gctgcattaa tgaatcggcc
     5581 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5641 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5701 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5761 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5821 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5881 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5941 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6001 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6061 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6121 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6181 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6241 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6301 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6361 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6421 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6481 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6541 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6601 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6661 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6721 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6781 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6841 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6901 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6961 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7021 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7081 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7141 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7201 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7261 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7321 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7381 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7441 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7501 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctg
//
