LOCUS       sequence-405329-        7186 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Zebrafish BMPR2a BMP receptor kinase domain and CTD fused to VfLOV.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7186)
  AUTHORS   Rogers KW, ElGamacy M, Jordan BM, Muller P
  TITLE     Optogenetic investigation of BMP target gene expression diversity.
  JOURNAL   Elife. 2020 Nov 11;9:e58641. doi: 10.7554/eLife.58641.
  PUBMED    33174840
REFERENCE   2  (bases 1 to 7186)
  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: "Elife."; 
            date: "2020-11-11"; volume: "9:e58641. doi"; pages: " 
            10.7554/eLife.58641"
COMMENT     Sequence Label: pCS2-Opto-BMPR2a
FEATURES             Location/Qualifiers
     source          1..7186
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      92..101
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             98..139
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSKPKDPSQR"
     CDS             3155..3181
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3235..3369
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3259..3278)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3313..3332
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3482..3502)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3482..3500)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3521..3537)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3521..3537)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3534..3556)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3545..3561
                     /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(3569..3575,3576..3593,3594..3599))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3614..3635
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3752..3769)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3923..4511)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4003..4022)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4682..5473,5474..5542))
                     /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     5305..5324
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5543..5647)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5673..6128)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5810..5831)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6022..6041
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6202..7186
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga tgccaccatg gggagtagca agagcaagcc
      121 taaggacccc agccagcgcg gtggaggagg ttctggaggc ggtggaagtg gtggcggagg
      181 tagccgcatg ctaagaggac gtggtaaaca ctcactgcat actttaaata tactggagac
      241 ggcactttct cctccttctt tggatctgga caacctcaca ctgcaggagc tgattggccg
      301 gggccggtac ggtacagtgt atcgtgcttc gctggatgac cgatctgtag cagtgaaagt
      361 tttcatttct gctaaccgcc agcagttcac taatgagcgt atgatttatc gcctcctcct
      421 ggatcacgag aacatagcgc gcttcctaga gagcgaggag cgcgtcggca cagaaggtcg
      481 gacagagttt ctccttctgc tggagtttta tcctcatggc tctctgtgca cgtatctgag
      541 cggccggact gtggactggt tgagctgctg ccgtttggct ctgtctgtga ccagagggtt
      601 ggcgtaccta catacagaga tacagcgagg ggatgtgtat aaaccggctg tctcccaccg
      661 ggatctgaac agcaggaatg tgctggttaa gacggacggc tcgtgtgtga tcagtgactt
      721 tggactgtcc atgattctga taggaaagag gccgcctggt catggagaag aggacaacag
      781 tgccatcagt gaggtaggta cagtgcggta catggctcca gaagtgctgg aaggggctgt
      841 gaatctgagg gattgtgagt ctgcgctgaa acaagtggat gtttatgcac tgggtctggt
      901 ttactgggag accttcatgc gctgtgcaga tcttttccca ggtgaaacag tgccagcttt
      961 tcagttggca ttccaggcag aggtgggcaa tcatcctacc atagaggacg tgcaggcgct
     1021 tgtgtccaga gaaaaagaaa gacccaaatt cccagaagcc tggaaagaga atagcctgac
     1081 ggtgcattct ctgaaagaga cgatggaaga ctgctgggat caggacgcag aagcccgact
     1141 gacagctcag tgtgcggagg agcgactcgc cgaactgctc ctcatctggg acagagaaaa
     1201 atcagccagt cctgctctta accacagcac tgcactgcag acacctaaag taggttctgt
     1261 ccttgaacac tcatccacaa acactgaaga ccatgaaggt gctgataaac cactccacaa
     1321 tgacacctca gtggagcaaa cctcagcagg aggaactaat cccgcagaga agaaaaagaa
     1381 ctgcatcaac tatgagtggc agcaggccca atcacgacag tctggcacag aaagctcctc
     1441 cgctactcct gtctcagagt cctgcaatgc cacgcatcct ccaccagtgg ccaccaatct
     1501 ctgtgctcag ctgacacgag aagacctgga gataccaaaa ctcgacccga gcgaagtcca
     1561 gaggaacatg agagagagct cggacgagag ccttatggaa cattcacaga aacagttctg
     1621 ctctcctgaa acgtcagcct cccacggccc gttttatcct ctcatgaaga tggtttctga
     1681 ggtttcgggg tcacaaggat ccagtaggca tggggacacc cctataacca tcttacctaa
     1741 gcagcagaac gtccctaaga gacccagtag cctcagtctc cacgtaaagc ctgggaaaac
     1801 atccacctcg acgtcttcat cgctgcggat gaagttcgga aagcttggaa agtcaaatct
     1861 gaagaaggtt gagatgggtg tggctaaaag cagtgtggtg aatgcgacgc atgaagcccg
     1921 gctgattaca gttgccaaca acgatgcggc ggcagcaatg aataaatatg caacagaacc
     1981 agcagccgag tcagcaggaa gttcagccaa cgaggacttg accttcggcc tcttaaacac
     2041 cagtcccgat gagcaggagc ctctgctgag aagagaggcg catcctgaca atgcaaacaa
     2101 caacaacagc aataacaaca atggtgaggg agatggtgat ggagagacag agggtggagg
     2161 agagggagga gagaacaatg agagtgtcgg tccgacaggg gatgcttcgt cgtcttctac
     2221 ggtggagccg gttgtgctgc ctggcccgtg ttctgcttca accgccattc ctccacaggc
     2281 ccaaagccag acacagacac acggagaggc tctgctcaga cagaaccgag tgcgcagacc
     2341 agagagaccc aactctctgg atctgtccat cacaacactg ccattactag gaggcaggtc
     2401 tgtcggcgat gggacagagg ggtcagggga taaaatcaag aagcgggtaa agacgcctta
     2461 cgcacttaag aagtggcgtc ctgccagctg ggttatcacc actgacacac tggatgccga
     2521 agtcaacaac aacagtcgtc acggaggagg cctcgggcag aatcagaatc aggctgggac
     2581 cagcagacct aaatcagctt cggctgtcta tctaggcagt cgaggaggat ctcgcttctc
     2641 agatcctaat gactgtgact ttggtggagg aggttctgga ggcggtggaa gtggtggcgg
     2701 aggtagccct gactacagtc tcgtgaaggc tctgcaaatg gcacaacaga attttgtcat
     2761 tacagacgcc tccctcccag acaaccctat cgtctacgcc agtagagggt ttctgacact
     2821 gacaggctat tctctcgacc agatcctggg caggaactgc aggtttctgc aagggccaga
     2881 aacagaccca agagctgtgg ataagatcag gaatgccatc accaaaggcg ttgataccag
     2941 tgtctgtctg ctgaattata gacaggatgg cacaaccttc tggaatctct tcttcgtggc
     3001 tggactcaga gattctaagg gcaatattgt caactacgtc ggagtgcagt caaaggtgag
     3061 cgaagattat gccaagctgc tggtcaacga gcagaacatt gagtacaaag gtgtgcgcac
     3121 cagtaacatg ctgcgcagaa agcccgggtc tagttatccg tacgacgtac cagactacgc
     3181 ataagaattc aaggcctctc gagcctctag aactatagtg agtcgtatta cgtagatcca
     3241 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     3301 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     3361 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     3421 gaggtttttt aattcgcggc cgcggcgcca atgcattggg cccggtaccc agcttttgtt
     3481 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
     3541 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     3601 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     3661 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3721 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3781 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3841 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3901 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3961 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4021 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4081 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4141 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4201 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4261 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4321 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     4381 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4441 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4501 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4561 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4621 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4681 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4741 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4801 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4861 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4921 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4981 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5041 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5101 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5161 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5221 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5281 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5341 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5401 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5461 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5521 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5581 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5641 ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa
     5701 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     5761 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     5821 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     5881 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     5941 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     6001 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     6061 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     6121 gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     6181 ctcttcgcta ttacgccagt cgaccatagc caattcaata tggcgtatat ggactcatgc
     6241 caattcaata tggtggatct ggacctgtgc caattcaata tggcgtatat ggactcgtgc
     6301 caattcaata tggtggatct ggaccccagc caattcaata tggcggactt ggcaccatgc
     6361 caattcaata tggcggactt ggcactgtgc caactgggga ggggtctact tggcacggtg
     6421 ccaagtttga ggaggggtct tggccctgtg ccaagtccgc catattgaat tggcatggtg
     6481 ccaataatgg cggccatatt ggctatatgc caggatcaat atataggcaa tatccaatat
     6541 ggccctatgc caatatggct attggccagg ttcaatacta tgtattggcc ctatgccata
     6601 tagtattcca tatatgggtt ttcctattga cgtagatagc ccctcccaat gggcggtccc
     6661 atataccata tatggggctt cctaataccg cccatagcca ctcccccatt gacgtcaatg
     6721 gtctctatat atggtctttc ctattgacgt catatgggcg gtcctattga cgtatatggc
     6781 gcctccccca ttgacgtcaa ttacggtaaa tggcccgcct ggctcaatgc ccattgacgt
     6841 caataggacc acccaccatt gacgtcaatg ggatggctca ttgcccattc atatccgttc
     6901 tcacgccccc tattgacgtc aatgacggta aatggcccac ttggcagtac atcaatatct
     6961 attaatagta acttggcaag tacattacta ttggaaggac gccagggtac attggcagta
     7021 ctcccattga cgtcaatggc ggtaaatggc ccgcgatggc tgccaagtac atccccattg
     7081 acgtcaatgg ggaggggcaa tgacgcaaat gggcgttcca ttgacgtaaa tgggcggtag
     7141 gcgtgcctaa tgggaggtct atataagcaa tgctcgttta gggaac
//
