LOCUS       sequence-338994-        6393 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses BMPR-2-Delta-Tail 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 6393)
  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 6393)
  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-Tail gRNA resistant
FEATURES             Location/Qualifiers
     source          1..6393
                     /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(3415..3434)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3480..3535
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3534..3553)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3896..3912)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3896..3912)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3909..3931)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3920..3936
                     /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(3944..3950,3951..3968,3969..3974))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3989..4010
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4069..4264
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4115..4250
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4270..4404
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4307..4326)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4361..4380
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4472..4489)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4643..5231)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4723..4742)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5402..6193,6194..6262))
                     /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     6025..6044
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6263..6367)
                     /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 ggctgcttct cagaatcaag aacggctgtg tgcatttaaa gatccatatc
     1861 aacaagatct tgggataggt gagagtcgaa tctctcatga aaatgggaca atattatgta
     1921 gtaagggatc aacatgctac ggtctgtggg agaaatcaaa aggggacatc aatcttgtga
     1981 aacaaggatg ttggtctcac atcggtgatc cccaagagtg ccactatgaa gagtgtgtag
     2041 taactaccac cccaccctca attcagaatg gaacgtaccg cttttgctgc tgtagtacag
     2101 atttatgtaa tgtcaacttt actgagaact ttccaccccc tgacacaaca ccactcagtc
     2161 cacctcattc atttaatcga gatgaaacga taatcattgc tttggcatca gtttctgtgt
     2221 tagctgtttt gatagtcgcc ttatgttttg gatacagaat gttgacagga gaccggaaac
     2281 agggtcttca cagcatgaac atgatggagg cggcagcagc agagccctcc cttgacctgg
     2341 ataacctgaa gctgctggag ctgattggac ggggtaggta tggtgctgtt tacaagggtt
     2401 ccttggatga gcgtccagtt gctgtaaaag tattttcttt tgcaaaccgt cagaatttta
     2461 taaatgaaaa aaacatttac agagtgcctt tgatggaaca tgacaacatt gctcgcttca
     2521 tagttggaga cgagaggctc actgcagacg gccgcatgga gtatttgctt gtgatggagt
     2581 attatcccaa tggatctctg tgcaaatatc tgagtctcca cacaagtgat tgggtaagct
     2641 cttgccgtct ggctcattct gtgactagag gactggctta tcttcacaca gaattaccac
     2701 gaggagatca ttataaaccc gcaatctccc accgagattt aaacagcagg aatgtcctgg
     2761 taaagaatga cggcgcgtgt gttatcagtg actttggttt atccatgagg ctaactggaa
     2821 atcggctggt gcgcccaggg gaagaagata atgcggctat aagtgaggtt ggcacaattc
     2881 gctatatggc accagaagtg ctagaaggag ctgtgaacct gagggactgt gagtcagctc
     2941 tgaagcaagt ggacatgtat gcgcttggac tcatctactg ggaggtgttt atgaggtgta
     3001 cagacctctt cccaggtgaa tctgtaccag attaccagat ggcttttcag acagaagttg
     3061 gaaaccatcc cacatttgag gatatgcagg ttcttgtgtc cagagagaag cagagaccca
     3121 agttcccaga agcctggaaa gaaaatagcc tggcagtgag gtcactcaag gaaacaattg
     3181 aagactgctg ggaccaggat gcagaggctc ggctcactgc acagtgtgct gaggagagga
     3241 tggctgaact catgatgata tgggagagaa acaagtctgt gagcccaacg gtcaacccaa
     3301 tgtcaactgc tatgcagaat gaacgctgag cggccgcact cctcaggtgc aggctgccta
     3361 tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact gagatctttt
     3421 tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta
     3481 ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg
     3541 aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt
     3601 ggcaacatat gcccatatgc tggctgccat gaacaaaggt tggctataaa gaggtcatca
     3661 gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg
     3721 ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc
     3781 cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt
     3841 ccctcttctc ttatggagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat
     3901 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     3961 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4021 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta
     4081 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4141 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4201 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4261 caaaaagcta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4321 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4381 aatgtatctt atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag
     4441 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4501 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4561 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4621 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4681 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4741 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4801 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4861 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4921 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4981 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5041 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5101 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5161 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5221 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5281 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5341 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5401 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5461 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5521 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5581 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5641 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5701 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5761 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5821 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5881 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5941 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6001 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6061 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6121 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6181 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6241 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6301 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6361 ttccgcgcac atttccccga aaagtgccac ctg
//
