LOCUS       sequence-191825-        7553 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Injection into Zebrafish larvae together with Tol2-Transposase to 
            achieve pan-neuronal expression of the photoconvertible fluorescent 
            protein Dendra2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7553)
  AUTHORS   Mohr MA, Argast P, Pantazis P
  TITLE     Labeling cellular structures in vivo using confined primed 
            conversion of photoconvertible fluorescent proteins.
  JOURNAL   Nat Protoc. 2016 Dec;11(12):2419-2431. doi: 10.1038/nprot.2016.134. 
            Epub 2016 Nov 3.
  PUBMED    27809312
REFERENCE   2  (bases 1 to 7553)
  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"; doi: 
            "10.1038/nprot.2016.134"; journalName: "Nat Protoc"; date: 
            "2016-12"; volume: "11"; issue: "12"; pages: "2419-2431"
COMMENT     Sequence Label: pMT-HuC:Dendra2
FEATURES             Location/Qualifiers
     source          1..7553
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(2635..2822,2823..2834)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(2868..2884)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2868..2884)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2921..2941)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2921..2939)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2960..2976)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2960..2976)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2973..2995)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2984..3000
                     /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(3008..3014,3015..3032,3033..3038))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3053..3074
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3191..3208)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3362..3950)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3442..3461)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4121..4912,4913..4981))
                     /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     4744..4763
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4982..5086)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5249..5270)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5461..5480
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5624..5646
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5638..5655
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5639..5655
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    join(5714..5725,5726..5863)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     polyA_signal    5955..6089
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5992..6011)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6046..6065
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        6096..6114
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(6260..6949)
                     /codon_start=1
                     /product="monomeric photoswitchable green-to-red 
                     fluorescent protein from Dendronephthya"
                     /label=Dendra2
                     /note="mammalian codon-optimized"
                     /translation="MNTPGINLIKEDMRVKVHMEGNVNGHAFVIEGEGKGKPYEGTQTA
                     NLTVKEGAPLPFSYDILTTAVHYGNRVFTKYPEDIPDYFKQSFPEGYSWERTMTFEDKG
                     ICTIRSDISLEGDCFFQNVRFKGTNFPPNGPVMQKKTLKWEPSTEKLHVRDGLLVGNIN
                     MALLLEGGGHYLCDFKTTYKAKKVVQLPDAHFVDHRIEILGNDSDYNKVKLYEHAVARY
                     SPLPSQVW"
     promoter        complement(6984..7002)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6985..7002)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 tccctccagc ccccaccatc tctccactct tccttttgcc ctccctttct tcctgccccc
       61 cccaaacaca catgcagagc tgagtgaata gagctccacc tgcactccga acagatgggt
      121 gcagaaaaag acaggagcca tgctttctcc ttgcccctcg ttctctctct ccctcatccc
      181 cttacttaaa ttttgtcctt tcttgatttc acattatttg tctttgtaga tcatttctaa
      241 gaatgcagtc cacgattaag tcctccattt gtgtggatgc agagcggctg taactgtgtg
      301 aaggtcatct ctggacaaaa caacctcgac ataaccgttt ctatattacc atgtgttgaa
      361 tgtgcgcatt acataatcga caaaggccaa cgctttaaaa tgacaactat ctgctcagct
      421 attacacgag gtgtggttat caagccggtt cgatgatact tgactggtaa gatgtgaaca
      481 tgaatgcacc catctcctta ttgttcccgg ttggtgctga tattaaacat cgacacagtg
      541 attgcagcgg ttgttatctc aagctccctt cccccatttc tagtctcagg gagtgggtag
      601 agctcaccga gacaggctgt ttggtctcat ctataatgca tctcctgccc caggacttga
      661 tgtgtgacaa atgacctatt tctgcacaga cccccacagg gccttgacct gagggagcgc
      721 gctctgtata tctttgttgt gcggatgctg gtaggtgcga gtctccataa gttggaagag
      781 atgcatgtag gcttatggaa atgcatgagg cgtgaggggg ttgttgttcc cagcggagga
      841 tgtgtttatt tcagtctgtt tatggagacc tcacctacat ctgcaccggc cttgggtgac
      901 cttgtggaaa cggttgtgtg gcgctcgtct tcaacacaac ttctcttgat tatgcagatt
      961 gtagacagtc accgcgcagg tatcaggcat caggagtcag cgaggtaccg agtgggtggt
     1021 attgtcgttg gcgggcaaat gtgctgtaat gcagatcaga aaaacagcat tttctcgtgt
     1081 ctgtgcataa tacacacaat tatatatatt ttgcttgagg agttcttggg aaaccatcag
     1141 agatgggcga gtgggcagag gcgttcaaac tcgttaacct ttgggtaatc cggagagctc
     1201 caatggttgc gcatttctgg ccaagtgttc tctggtaatc ttgacacatc ctgcccctct
     1261 gtcctctcct ccctttagtg gtcactttca ctttcagttc ttgctctcgg ttctcgtccc
     1321 gcgtctctac cccagtcaga ttctgagtgt gggccctgtc tgatatttta cataattgcc
     1381 ttacataagt ggccagctca gctcagagct tggatgatat tagtggcgat gaatattgca
     1441 tgaggaaggc actcgttaag cgttaataag aggatggaga tgggtcccgc tgtcagtctg
     1501 tcttaacccc gcccactctg ctctgtcaac tcgccacgtt tcttagaccc agccgagggc
     1561 gccattatag gtgcgggtgt tgttttctgc tttgtgagcc gaataaaggc agcacttgtg
     1621 gcacaaagaa cccaggtctg ttagctgccc acagttccgc acgcaccccg ttcacactta
     1681 acatgaataa acttaattgc agaggctcat tagggggtct ccgctggtag cggggcagct
     1741 gaatccagcc cgtgggtaag agggcatctg aagcacctat tcaccgcccc gacgcagagg
     1801 atcaagcatg ccagttgctt gaagctcttt cggtaatttc gctgataatt tttaatgttc
     1861 aggagcccct ccagaagagc ccattaagag actgtcactt ttgtccccag tggcattaca
     1921 gcgtaagagt ctctcttgtg ctctttcaat gatcattgat tttagacact ttttatggcc
     1981 ttccaaacgc tgtggggatg actggtggat gaaactaagc aatatactct tgttttttta
     2041 tccaatatct caaaagtgtc tgttttacat gtgcaaattg taaatttaca tttatgagtg
     2101 gaaagcatgt tatgatcacg ctgaatctgc ttagcaccgg cgcaggagat atgattgtgt
     2161 gaaacagatg tgttccattg ctacagctag tgttatccct tcttattatg gtctgaggta
     2221 acggattagg atggctcaaa tgactgcgct aaagacgatt ttttccagga tttgtatttt
     2281 caattggcag tgttttaaaa tggcttaatt tggataaagc ttgtaatgtt atttcataca
     2341 tcaaaatgtg ctcagtagga tgtatacaac taaatatgtt tactcgtaaa catcactcta
     2401 tacatcatca cttgcagtca gtatatggtt ttagagttca taacataaat gagtacacca
     2461 gctttttaaa aattgatatt tttataaatt tcactataca tatggttctt tgaaacaaaa
     2521 aagttttaat aaacagtttt atttattaaa ataagatttt ggtcacccaa ggtctttagg
     2581 aatagaaaga taatgcattt aaattcaaat tagtgaattc ctcgagccgg gcccaagtga
     2641 tctccaaaaa ataagtactt tttgactgta aataaaattg taaggagtaa aaagtacttt
     2701 tttttctaaa aaaatgtaat taagtaaaag taaaagtatt gatttttaat tgtactcaag
     2761 taaagtaaaa atccccaaaa ataatactta agtacagtaa tcaagtaaaa ttactcaagt
     2821 actttacacc tctggttctt gaccccctac cttcagcaag cccagcagat ccactagttc
     2881 tagagcggcc gccaccgcgg tggagctcca gcttttgttc cctttagtga gggttaattg
     2941 cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     3001 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     3061 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     3121 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     3181 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3241 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3301 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3361 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3421 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3481 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3541 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3601 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3661 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3721 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3781 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3841 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3901 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3961 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4021 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4081 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4141 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4201 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4261 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4321 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4381 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4441 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4501 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4561 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4621 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4681 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4741 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4801 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4861 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4921 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4981 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5041 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5101 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     5161 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     5221 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     5281 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     5341 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     5401 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     5461 agggaagaaa gcgaaaggag cgggcgctag ggcgcgtccc attcgccatt caggctgcgc
     5521 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     5581 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     5641 aaaacgacgg ccagtgagcg cgcgtaatac gacgtcctat agggcgaatt gggtaccggc
     5701 atatggttct tgacagaggt gtaaaaagta ctcaaaaatt ttactcaagt gaaagtacaa
     5761 gtacttaggg aaaattttac tcaattaaaa gtaaaagtat ctggctagaa tcttacttga
     5821 gtaaaagtaa aaaagtactc cattaaaatt gtacttgagt attagatctg catgcgttta
     5881 aacccgggat atcaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     5941 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     6001 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     6061 tcatcaatgt atcttatcat gtctggatca ccggttaata cgactcacta tagggagtcc
     6121 ggatacgtaa tacgactcac tatagttcta gaggctcgag cggccgccag tgtgatggat
     6181 atctgcagaa ttcgcccttt caacaggagt ccaagctcag ctaattaagc tttcacttgt
     6241 acacgccgct gtcgccggac cacacctggc tgggcagggg gctgtagcgg gccacggcgt
     6301 gctcgtacag cttcaccttg ttgtagtcgc tgtcgttgcc caggatctcg atgcggtggt
     6361 ccacgaagtg ggcgtcgggc agctgcacca ccttcttggc cttgtaggtg gtcttgaagt
     6421 cgcacaggta gtggccgccg ccctccagca gcagggccat gttgatgttg cccaccagca
     6481 ggccgtcgcg cacgtgcagc ttctcggtgc tgggctccca cttcagggtc ttcttctgca
     6541 tcacggggcc gttggggggg aagttggtgc ccttgaagcg cacgttctgg aagaagcagt
     6601 cgccctccag gctgatgtcg ctgcggatgg tgcagatgcc cttgtcctcg aaggtcatgg
     6661 tgcgctccca gctgtagccc tcggggaagc tctgcttgaa gtagtcgggg atgtcctcgg
     6721 ggtacttggt gaacacccgg ttgccgtagt gcacggcggt ggtcaggatg tcgtagctga
     6781 agggcagggg ggcgccctcc ttcacggtca ggttggcggt ctgggtgccc tcgtagggct
     6841 tgcccttgcc ctcgccctcg atcacgaagg cgtggccgtt cacgttgccc tccatgtgca
     6901 ccttcacgcg catgtcctcc ttgatcaggt taattcccgg ggtgttcatg gatcctgcaa
     6961 aaagaacaag tagcttgtac tcttctatag tgtcacctaa ataggcctag gtcttgacgt
     7021 acaaagatga tattgatcta ggtccttcga tttgcaggtc ttcaggctgg attgcagata
     7081 ttattacgcc gtggcttgga atggagaaat gcaaatgtca gtttccaaag gaaaagaaaa
     7141 aaaaagaaaa aggaaaaagg acagtcttcg ccaggaaatc tcagcccgtg cgattgtagg
     7201 ttttcctcag cagtgtgaaa atgaggataa gggggttttc ttcaggatga tgatgagaga
     7261 gagatttctc tccgtgtggc gtgggcgatg gttcagaaaa aaagaaaagc agtgatcccc
     7321 agtagggtgt gtccagggga gggtgttgtg gttctgctgt tgtgtgtgta tttcccagct
     7381 cctcgcaggc tccttctgtg cctctgctgc aatgttcttc ttttatgtgt ccttctggct
     7441 ggcagagctt gctgctgatt ggctaattgt gcagggaggg tactacatgg cagccattgc
     7501 aaagcgctgc tctcccatta tccccttcag gagagatcac cgagaacccc cca
//
