LOCUS       sequence-338987-        7022 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses BMPR-2-Delta-Kinase 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 7022)
  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 7022)
  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-Kinase gRNA resistant
FEATURES             Location/Qualifiers
     source          1..7022
                     /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(4044..4063)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4109..4164
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4163..4182)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4525..4541)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4525..4541)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4538..4560)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4549..4565
                     /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(4573..4579,4580..4597,4598..4603))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4618..4639
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4698..4893
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4744..4879
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4899..5033
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4936..4955)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4990..5009
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5101..5118)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5272..5860)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5352..5371)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6031..6822,6823..6891))
                     /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     6654..6673
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6892..6996)
                     /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 gccrggsggg 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 tatggctgaa ctcatgatga tatgggagag aaacaagtct gtgagcccaa cggtcaaccc
     2401 aatgtcaact gctatgcaga atgaacgcaa cctgtcacat aataggcgtg tgccaaaaat
     2461 cgggccttac ccagattatt cctcttcctc atatattgaa gactctatcc atcatactga
     2521 cagcattgtg aagaatattt cctctgagca ttcgatgtcc agcacaccat tgacaatagg
     2581 agaaaagaat cgaaattcaa ttaattatga acgacagcaa gcacaagctc gaatccctag
     2641 cccagaaaca agcgtcacaa gcctgtccac aaacacaacc accacaaaca ccaccggcct
     2701 cactccaagt actggcatga ccactatatc tgagatgcca tacccagatg agacacattt
     2761 gcacgccaca aatgttgcac agtcaatcgg gccaacccct gtctgcttac agctgacaga
     2821 agaagacttg gagactaata agctagatcc aaaagaagtt gataagaacc tcaaggaaag
     2881 ctctgatgag aatctcatgg agcattctct gaagcagttc agtgggccag acccattgag
     2941 cagtaccagt tctagcttgc tttatccact cataaagctc gcagtggaag tgactggaca
     3001 acaggacttc acacaggctg caaatgggca agcatgttta attcctgatg ttccacctgc
     3061 tcagatctat cctctcccta agcaacagaa ccttcctaag agacctacta gtttgccttt
     3121 gaacaccaaa aattcaacaa aagaaccccg gctaaaattt ggcaacaagc acaaatcaaa
     3181 cttgaaacaa gtagaaactg gagttgccaa gatgaataca atcaatgcag cagagcctca
     3241 tgtggtgaca gtaactatga atggtgtggc aggtagaagc cacaatgtta attcccatgc
     3301 tgccacaacc cagtatgcca atggcgcagt gccagctggc caggcagcca acatagtggc
     3361 acataggtcc caagaaatgc tgcagaatca atttattggt gaggatacca ggctgaatat
     3421 caattccagt cctgatgagc atgaaccttt actgagacga gagcaacagg ctggccatga
     3481 tgaaggggtt ctggatcgtt tggtagatag gagggaacgg ccattagaag gtggccgaac
     3541 aaattccaat aacaacaaca gcaatccatg ttcagaacaa gatatcctta cacaaggtgt
     3601 tacaagcaca gctgcagatc ctgggccatc aaagcccaga agagcacaga ggcccaattc
     3661 tctggatctt tcagccacaa atatcctgga tggcagcagt atacagatag gtgagtcaac
     3721 acaagatggc aaatcaggat caggtgaaaa gatcaagaga cgtgtgaaaa ctccatactc
     3781 tcttaagcgg tggcgcccgt ccacctgggt catctccacc gagccgctgg actgtgaggt
     3841 caacaacaat ggcagtgaca gggcagtcca ttctaaatct agcactgctg tgtaccttgc
     3901 agagggaggc actgccacga ccacagtgtc taaagatata ggaatgaatt gtctgtgagc
     3961 ggccgcactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc
     4021 ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg ggacatcatg
     4081 aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg
     4141 tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa
     4201 catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct ggctgccatg
     4261 aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc
     4321 ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta
     4381 tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc
     4441 ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc cctcgacctg
     4501 cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4561 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4621 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4681 gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4741 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4801 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     4861 ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt gcagcttata
     4921 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4981 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccgctgca
     5041 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5101 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5161 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5221 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5281 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5341 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5401 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5461 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5521 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5581 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5641 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5701 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5761 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5821 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5881 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5941 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6001 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6061 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6121 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6181 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6241 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6301 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6361 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6421 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6481 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6541 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6601 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6661 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6721 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6781 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6841 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6901 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6961 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7021 tg
//
