LOCUS       sequence-314977-        5148 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Amyloid Beta-mCherry in Zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5148)
  AUTHORS   Kaur P, Kibat C, Teo E, Gruber J, Mathuru A, Tolwinski ANS
  TITLE     Use of Optogenetic Amyloid-beta to Monitor Protein Aggregation in 
            Drosophila melanogaster, Danio rerio and Caenorhabditis elegans.
  JOURNAL   Bio Protoc. 2020 Dec 5;10(23):e3856. doi: 10.21769/BioProtoc.3856. 
            eCollection 2020 Dec 5.
  PUBMED    33659494
REFERENCE   2  (bases 1 to 5148)
  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: "Bio 
            Protoc."; date: "2020-12-5"; volume: "10(23):e3856. doi"; pages: " 
            10.21769/BioProtoc.3856. eCollection 2020 Dec 5"
COMMENT     Sequence Label: pDEST-Tol2-PA2-CMV-AB-mCh
FEATURES             Location/Qualifiers
     source          1..5148
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      334..343
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             340..1044
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELY"
     primer_bind     complement(485..503)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     753..772
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1050..1184
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1074..1093)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1128..1147
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(1623..1810,1811..1822)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(1856..1872)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1856..1872)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1909..1929)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1909..1927)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1948..1964)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1948..1964)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1961..1983)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1972..1988
                     /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(1996..2002,2003..2020,2021..2026))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2041..2062
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2179..2196)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2350..2938)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2430..2449)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3109..3900,3901..3969))
                     /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     3732..3751
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3970..4074)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4237..4258)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(4607..4618,4619..4781)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctatggat gcagaattcc gacatgactc aggatatgaa
      241 gttcatcatc aaaaattggt gttctttgca gaagatgtgg gttcaaacaa aggtgcaatc
      301 attggactca tggtgggcgg tgttgtcata gcggccacca tggtgagcaa gggcgaggag
      361 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
      421 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
      481 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
      541 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
      601 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
      661 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
      721 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
      781 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
      841 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
      901 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
      961 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     1021 accggcggca tggacgagct gtacatgatg atccagacat gataagatac attgatgagt
     1081 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     1141 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     1201 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     1261 tctacaaatg tggtatggct gattatgatc ctctagatca gatctgcgaa gatacggcca
     1321 cgggtgctct tgatcctgtg gctgattttg gactgtgctg ctcgcagctg ctgatgaatc
     1381 acatacttcc tccattttct tccactgatt gactgttata atttccctaa tttccaggtc
     1441 aaggtgctgt gcattgtggt aatagatgtg acatgacgtc acttccaaag gaccaatgaa
     1501 catgtctgac caatttcata taatgtgaaa acgattttca taggcagaat aaataacatt
     1561 taaattaaac tgggcatcag cgcaattcaa ttggtttggt aatagcaagg gaaaatagaa
     1621 tgaagtgatc tccaaaaaat aagtactttt tgactgtaaa taaaattgta aggagtaaaa
     1681 agtacttttt tttctaaaaa aatgtaatta agtaaaagta aaagtattga tttttaattg
     1741 tactcaagta aagtaaaaat ccccaaaaat aatacttaag tacagtaatc aagtaaaatt
     1801 actcaagtac tttacacctc tggttcttga ccccctacct tcagcaagcc cagcagatcc
     1861 actagttcta gagcggccgc caccgcggtg gagctccagc ttttgttccc tttagtgagg
     1921 gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     1981 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     2041 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     2101 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2161 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     2221 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2281 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2341 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2401 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2461 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2521 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2581 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2641 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2701 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2761 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2821 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2881 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2941 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3001 cattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3061 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3121 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3181 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3241 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3301 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     3361 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     3421 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     3481 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     3541 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     3601 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     3661 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     3721 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     3781 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     3841 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     3901 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     3961 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4021 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4081 tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     4141 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     4201 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     4261 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     4321 cgctggctaa gaactcatca gcctccccgg tccatctacc cacgtaccaa tgcaccaatt
     4381 ggccacaatg acggctacta catggtgcca ttccttcctc tttataggaa tggagactac
     4441 ctcctgtcca acaatgctct tggatacgag tacgcctacc tgttggaccc aggtcattgc
     4501 acaacaccag aaatgccctc tgatctgcaa aagacgtgaa tatctgttca gacacccata
     4561 tccactctgt tccacacagg tcagaggttt gtccaggagt tcttgacaga ggtgtaaaaa
     4621 gtactcaaaa attttactca agtgaaagta caagtactta gggaaaattt tactcaatta
     4681 aaagtaaaag tatctggcta gaatcttact tgagtaaaag taaaaaagta ctccattaaa
     4741 attgtacttg agtattaagg aagtaaaagt aaaagcaaga aagaaaacta gagattcttg
     4801 tttaagcttt taatctcaaa aaacattaaa tgaaatgcat acaaggtttt atcctgcttt
     4861 agaactgttt gtatttaatt atcaaactat aagacagaca atctaatgcc agtacacgct
     4921 actcaaagtt gtaaaacctc agatttaact tcagtagaag ctgattctca aaattgttag
     4981 tgtcaagcct agctcttttg gggctgaaaa gcaatcctgc agtgctgaaa agcctctcac
     5041 aggcagccga tgcgggaaga ggtgtattag tcttgataga gaggctgcaa atagcaggaa
     5101 acgtgagcag agactccctg gtgtctgaaa cacaggccag atgggccc
//
