LOCUS       sequence-409800-        5757 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Zebrafish Alk3 BMP receptor kinase domain fused to VfLOV.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5757)
  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 5757)
  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-Alk3
FEATURES             Location/Qualifiers
     source          1..5757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        22..999
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1034..1052
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1034..1051
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1084..1125
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSKPKDPSQR"
     CDS             2728..2754
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2808..2942
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2832..2851)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2886..2905
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3055..3075)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3055..3073)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3094..3110)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3094..3110)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3107..3129)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3118..3134
                     /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(3142..3148,3149..3166,3167..3172))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3187..3208
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3325..3342)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3496..4084)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3576..3595)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4255..5046,5047..5115))
                     /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     4878..4897
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5116..5220)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5246..5701)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5383..5404)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5595..5614
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tcgctattac gccagtcgac cgccaattca atatggcgta tatggactca tgccaattca
       61 atatggtgga tctggacctg tgccaattca atatggcgta tatggactcg tgccaattca
      121 atatggtgga tctggacccc agccaattca atatggcgga cttggcacca tgccaattca
      181 atatggcgga cctggcactg tgccaactgg ggaggggtct acttggcacg gtgccaagtt
      241 tgaggagggg tcttggccct gtgccaagtc cgccatattg aattggcatg gtgccaataa
      301 tggcggccat attggctata tgccaggatc aatatatagg caatatccaa tatggcccta
      361 tgccaatatg gctattggcc aggttcaata ctatgtattg gccctatgcc atatagtatt
      421 ccatatatgg gttttcctat tgacgtagat agcccctccc aatgggcggt cccatatacc
      481 atatatgggg cttcctaata ccgcccatag ccactccccc attgacgtca atggtctcta
      541 tatatggtct ttcctattga cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc
      601 ccattgacgt caattacggt aaatggcccg cctggctcaa tgcccattga cgtcaatagg
      661 accacccacc attgacgtca atgggatggc tcattgccca ttcatatccg ttctcacgcc
      721 ccctattgac gtcaatgacg gtaaatggcc cacttggcag tacatcaata tctattaata
      781 gtaacttggc aagtacatta ctattggaag tacgccaggg tacattggca gtactcccat
      841 tgacgtcaat ggcggtaaat ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa
      901 tggggagggg caatgacgca aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc
      961 taatgggagg tctatataag caatgctcgt ttagggaacc gccattctgc ctggggacgt
     1021 cggagcaagc ttgatttagg tgacactata gaatacaagc tacttgttct ttttgcagga
     1081 tccatgggga gtagcaagag caagcctaag gaccccagcc agcgcggtgg aggaggttct
     1141 ggaggcggtg gaagtggtgg cggaggtagc tacaggtata agtggcagac agagaggcag
     1201 cgctaccaca gagacctgga gcaagacgag gcctttatcc cagcaggaga atccctgaaa
     1261 gacctcatca accagtctca gacctcaggc agtggctctg gactccctct gctggtgcag
     1321 cgcactatag caaagcagat ccagacagtg cgaatgatcg gaaaaggacg atatggagaa
     1381 gtgtggcttg gtcggtggag aggagagaag gtagcagtga aggtgttctt tacccgagag
     1441 gaggccagct ggttcagaga gacagagatc tatcaaaccg tgctcatgag acatgagaac
     1501 atactcggct ttatcgctgc tgatataaat ggcacaggag cctctacgca gctgtacctg
     1561 atcacagact accatgagaa tggctctctg tatgactatc tgaagttcac gactttggac
     1621 acacaggctc tactcagact ggccttctct gcagcctgtg gcctgtgtca cctgcatacg
     1681 gagatctacg gcacgcaggg aaaaccagcg atcgctcaca gagacctgaa gagcaagaac
     1741 attctcatca agaaaaacgg cacctgctgc atcgctgacc tcggccttgc tgtgaaattc
     1801 aacagtgaca caaatgaagt ggacctccca ttaagcacac gtatgggaac caggcgctac
     1861 atggctccag aagtgttgga cgagactctg aataagaatc atttccaggc ctacatcatg
     1921 gcagacatct acagctatgg gctggttatt tgggaaatgg ccagacgctg tgtcactgga
     1981 gggattgttg aggagtatca gctgccatat tatgagatgg tgccttcaga cccatcttat
     2041 gaagacatgt tggaggttgt ttgtgtcaaa ggactgcggc ccaccgtatc caacagatgg
     2101 aacagtgatg agtgcttaag ggccatgcta aagctgatgt ctgaatgctg ggcccacaat
     2161 cctgcatcac gcttaaccat cctacgagtc aagaagactt tagccaaaat ggtggaatct
     2221 caagacatta aaatcggtgg aggaggttct ggaggcggtg gaagtggtgg cggaggtagc
     2281 cctgactaca gtctcgtgaa ggctctgcaa atggcacaac agaattttgt cattacagac
     2341 gcctccctcc cagacaaccc tatcgtctac gccagtagag ggtttctgac actgacaggc
     2401 tattctctcg accagatcct gggcaggaac tgcaggtttc tgcaagggcc agaaacagac
     2461 ccaagagctg tggataagat caggaatgcc atcaccaaag gcgttgatac cagtgtctgt
     2521 ctgctgaatt atagacagga tggcacaacc ttctggaatc tcttcttcgt ggctggactc
     2581 agagattcta agggcaatat tgtcaactac gtcggagtgc agtcaaaggt gagcgaagat
     2641 tatgccaagc tgctggtcaa cgagcagaac attgagtaca aaggtgtgcg caccagtaac
     2701 atgctgcgca gaaagcccgg gtctagttat ccgtacgacg taccagacta cgcataagaa
     2761 ttcaaggcct ctcgagcctc tagaactata gtgagtcgta ttacgtagat ccagacatga
     2821 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     2881 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     2941 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     3001 tttaattcgc ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta
     3061 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     3121 ttatccgctc acaattccac acaacatacg agccgggagc ataaagtgta aagcctgggg
     3181 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3241 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3301 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3361 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3421 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3481 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3541 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3601 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3661 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3721 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3781 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3841 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3901 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3961 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4021 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4081 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4141 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4201 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4261 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4321 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4381 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4441 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4501 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4561 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4621 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4681 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4741 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4801 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4861 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4921 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4981 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5041 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5101 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5161 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5221 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
     5281 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     5341 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     5401 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     5461 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     5521 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     5581 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     5641 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     5701 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctct
//
