LOCUS       sequence-409801-        5781 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Zebrafish Alk8 BMP receptor kinase domain fused to VfLOV.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5781)
  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 5781)
  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-Alk8
FEATURES             Location/Qualifiers
     source          1..5781
                     /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"
     CDS             85..126
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSKPKDPSQR"
     CDS             1753..1779
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1833..1967
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1857..1876)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1911..1930
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2080..2100)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2080..2098)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2119..2135)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2119..2135)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2132..2154)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2143..2159
                     /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(2167..2173,2174..2191,2192..2197))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2212..2233
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2350..2367)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2521..3109)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2601..2620)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3280..4071,4072..4140))
                     /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     3903..3922
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4141..4245)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4271..4726)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4408..4429)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4620..4639
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4797..5781
                     /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 atccatgggg agtagcaaga gcaagcctaa ggaccccagc
      121 cagcgcggtg gaggaggttc tggaggcggt ggaagtggtg gcggaggtag ccgtcgactc
      181 catcatgggc gtctggagag actgcacgag tttgacactg aacagggggc catcgatggg
      241 cttatcgcgt ctaatgtcgg agacagcaca cttgcggatc tgatggatca ctcctgcact
      301 tcaggcagtg gttcaggact gcccttcctg gttcagagaa cggttgcgcg gcagatcagc
      361 ctggtggagt gtgttggtaa aggacggtac ggtgaagtgt ggagaggtca atggcaagga
      421 gaaaatgtag ccgtgaagat cttttcctct agagatgaga agtcatggtt tcgagaaaca
      481 gaaatttaca acactgttct gctacgacat gaaaatatat taggcttcat ggcttctgac
      541 atgacctccc gaaactctag cactcagctg tggctgatca cacactatca cgagaatggc
      601 tctctgtatg actacctgca gcgtgtggct gtggagatgg cagatggact gcacatggcg
      661 gcttcgattg ccagcgggct ggtgcacctg cacacggaga tctttggcac ggagggcaaa
      721 ccggccatcg ctcacagaga cctgaagagc aagaacatcc ttgtgaagaa agatttgcag
      781 tgctgcatcg ctgacctggg tctggcagta acacacacgc agtctgataa tcagcttgat
      841 gtgggaaata atcctaaagt gggaaccaaa cgctacatgg caccggaggt tctagatgag
      901 accattcaga cggactgttt tgacgcctat aagagggtgg atatctgggc ctttgggttg
      961 gtgctgtggg agatcgcacg cagaaccatc agcaatggaa ttgtagagga atacaagccg
     1021 cctttctatg acctggttcc taatgatccc agctttgacg acatgaggaa agtggtttgt
     1081 gtggagcagc aaaggccatt cattcccaac cgctggtttt cagatcctac cctgtctgct
     1141 ctggtgaagc tgatgaaaga gtgctggtac cagaacccct cggctcgtct cactgccctg
     1201 cgcatcaaaa agactctgga taaaatccac agttcactgg agaagggcaa aaccgactgc
     1261 ggtggaggag gttctggagg cggtggaagt ggtggcggag gtagccctga ctacagtctc
     1321 gtgaaggctc tgcaaatggc acaacagaat tttgtcatta cagacgcctc cctcccagac
     1381 aaccctatcg tctacgccag tagagggttt ctgacactga caggctattc tctcgaccag
     1441 atcctgggca ggaactgcag gtttctgcaa gggccagaaa cagacccaag agctgtggat
     1501 aagatcagga atgccatcac caaaggcgtt gataccagtg tctgtctgct gaattataga
     1561 caggatggca caaccttctg gaatctcttc ttcgtggctg gactcagaga ttctaagggc
     1621 aatattgtca actacgtcgg agtgcagtca aaggtgagcg aagattatgc caagctgctg
     1681 gtcaacgagc agaacattga gtacaaaggt gtgcgcacca gtaacatgct gcgcagaaag
     1741 cccgggtcta gttatccgta cgacgtacca gactacgcat aagaattcaa ggcctctcga
     1801 gcctctagaa ctatagtgag tcgtattacg tagatccaga catgataaga tacattgatg
     1861 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     1921 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     1981 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa ttcgcggccg
     2041 cggcgccaat gcattgggcc cggtacccag cttttgttcc ctttagtgag ggttaattgc
     2101 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     2161 tccacacaac atacgagccg ggagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     2221 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     2281 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     2341 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2401 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2461 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2521 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2581 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2641 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2701 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2761 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2821 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2881 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2941 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3001 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3061 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3121 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3181 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3241 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3301 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3361 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3421 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3481 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3541 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3601 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3661 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3721 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3781 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3841 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     3901 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3961 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4021 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4081 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4141 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4201 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4261 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     4321 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     4381 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     4441 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     4501 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     4561 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     4621 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     4681 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
     4741 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagtcg
     4801 accgccaatt caatatggcg tatatggact catgccaatt caatatggtg gatctggacc
     4861 tgtgccaatt caatatggcg tatatggact cgtgccaatt caatatggtg gatctggacc
     4921 ccagccaatt caatatggcg gacttggcac catgccaatt caatatggcg gacctggcac
     4981 tgtgccaact ggggaggggt ctacttggca cggtgccaag tttgaggagg ggtcttggcc
     5041 ctgtgccaag tccgccatat tgaattggca tggtgccaat aatggcggcc atattggcta
     5101 tatgccagga tcaatatata ggcaatatcc aatatggccc tatgccaata tggctattgg
     5161 ccaggttcaa tactatgtat tggccctatg ccatatagta ttccatatat gggttttcct
     5221 attgacgtag atagcccctc ccaatgggcg gtcccatata ccatatatgg ggcttcctaa
     5281 taccgcccat agccactccc ccattgacgt caatggtctc tatatatggt ctttcctatt
     5341 gacgtcatat gggcggtcct attgacgtat atggcgcctc ccccattgac gtcaattacg
     5401 gtaaatggcc cgcctggctc aatgcccatt gacgtcaata ggaccaccca ccattgacgt
     5461 caatgggatg gctcattgcc cattcatatc cgttctcacg ccccctattg acgtcaatga
     5521 cggtaaatgg cccacttggc agtacatcaa tatctattaa tagtaacttg gcaagtacat
     5581 tactattgga agtacgccag ggtacattgg cagtactccc attgacgtca atggcggtaa
     5641 atggcccgcg atggctgcca agtacatccc cattgacgtc aatggggagg ggcaatgacg
     5701 caaatgggcg ttccattgac gtaaatgggc ggtaggcgtg cctaatggga ggtctatata
     5761 agcaatgctc gtttagggaa c
//
