LOCUS Exported 7022 bp ds-DNA circular SYN 04-AUG-2021 DEFINITION Expresses BMPR-2-Delta-Kinase resistant to the gRNAs(#163388-90) in mammalian cells. ACCESSION . VERSION . KEYWORDS pCAG-Bmpr2-Delta-Kinase gRNA resistant 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 Aug 4, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7022 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 5..384 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" 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(4573..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" primer_bind 4806..4825 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" 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(6031..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 //