LOCUS       sequence-340821-        6003 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses BMP-8A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6003)
  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 6003)
  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-Bmp8a
FEATURES             Location/Qualifiers
     source          1..6003
                     /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"
     regulatory      1734..1743
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3025..3044)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3090..3145
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3144..3163)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3506..3522)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3506..3522)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3519..3541)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3530..3546
                     /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(3554..3560,3561..3578,3579..3584))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3599..3620
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3679..3874
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3725..3860
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3880..4014
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3917..3936)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3971..3990
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4082..4099)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4253..4841)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4333..4352)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5012..5803,5804..5872))
                     /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     5635..5654
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5873..5977)
                     /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 tggctatgcg tcccgggcca ctctggctat tgggccttgc tctgtgcgcg ctgggaggcg
     1801 gccacggtcc gcgtcccccg cacacctgtc cccagcgtcg cctgggagcg cgcgagcgcc
     1861 gcgacatgca gcgtgaaatc ctggcggtgc tcgggctacc gggacggccc cgaccccgtg
     1921 cacaacccgc cgctgcccgg cagccagcgt ccgcgcccct cttcatgttg gacctatacc
     1981 acgccatgac cgatgacgac gacggcgggc caccacaggc tcacttaggc cgtgccgacc
     2041 tggtcatgag cttcgtcaac atggtggaac gcgaccgtac cctgggctac caggagccac
     2101 actggaagga attccacttt gacctaaccc agatccctgc tggggaggct gtcacagctg
     2161 ctgagttccg gatctacaaa gaacccagca cccacccgct caacacaacc ctccacatca
     2221 gcatgttcga agtggtccaa gagcactcca acagggagtc tgacttgttc tttttggatc
     2281 ttcagacgct ccgatctggg gacgagggct ggctggtgct ggacatcaca gcagccagtg
     2341 accgatggct gctgaaccat cacaaggacc tgggactccg cctctatgtg gaaaccgcgg
     2401 atgggcacag catggatcct ggcctggctg gtctgcttgg acgacaagca ccacgctcca
     2461 gacagccttt catggtaacc ttcttcaggg ccagccagag tcctgtgcgg gcccctcggg
     2521 cagcgagacc actgaagagg aggcagccaa agaaaacgaa cgagcttccg caccccaaca
     2581 aactcccagg gatctttgat gatggccacg gttcccgcgg cagagaggtt tgccgcaggc
     2641 atgagctcta cgtcagcttc cgtgaccttg gctggctgga ctgggtcatc gccccccagg
     2701 gctactctgc ctattactgt gagggggagt gtgctttccc actggactcc tgtatgaacg
     2761 ccaccaacca tgccatcttg cagtctctgg tgcacctgat gaagccagat gttgtcccca
     2821 aggcatgctg tgcacccacc aaactgagtg ccacctctgt gctgtactat gacagcagca
     2881 acaatgtcat cctgcgtaaa caccgtaaca tggtggtcaa ggcctgtggc tgccactgag
     2941 cggccgcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc
     3001 cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat
     3061 gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt
     3121 gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa
     3181 acatcagaat gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat
     3241 gaacaaaggt tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc
     3301 cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt
     3361 atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct
     3421 cctctcctga ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacct
     3481 gcagcccaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     3541 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     3601 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3661 tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3721 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3781 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     3841 aggaggcttt tttggaggcc taggcttttg caaaaagcta acttgtttat tgcagcttat
     3901 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     3961 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatccgctgc
     4021 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4081 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4141 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4201 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4261 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4321 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4381 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4441 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4501 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4561 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4621 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4681 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4741 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4801 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4861 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4921 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4981 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5041 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5101 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5161 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5221 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5281 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5341 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5401 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5461 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5521 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5581 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5641 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5701 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5761 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5821 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5881 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5941 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6001 ctg
//
