LOCUS       sequence-369602-        5574 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of a fusion of the transmembrane protein TVA and the red 
            fluorescent protein mCherry under UAS control..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5574)
  AUTHORS   Satou C, Neve RL, Oyibo HK, Zmarz P, Huang KH, Arn Bouldoires E, 
            Mori T, Higashijima SI, Keller GB, Friedrich RW
  TITLE     A viral toolbox for conditional and transneuronal gene expression in
            zebrafish.
  JOURNAL   Elife. 2022 Jul 22;11. pii: 77153. doi: 10.7554/eLife.77153.
  PUBMED    35866706
REFERENCE   2  (bases 1 to 5574)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2022-07-22"; volume: "11. pii: 77153. doi"; pages: " 
            10.7554/eLife.77153"
COMMENT     Sequence Label: UAS:TVA-mCherry
FEATURES             Location/Qualifiers
     source          1..5574
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    593..687
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     CDS             1161..1868
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1303..1321)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1571..1590
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1891..1908)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1897..2008
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2728..2748)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2728..2746)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2767..2783)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2767..2783)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2780..2802)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2791..2807
                     /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(2815..2821,2822..2839,2840..2845))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2860..2881
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2998..3015)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3169..3757)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3249..3268)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3928..4719,4720..4788))
                     /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     4551..4570
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4789..4893)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4919..5374)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5056..5077)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5268..5287
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5501..5523
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5515..5532
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5516..5532
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5542..5561
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5542..5560
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggcatatggt tcttgacaga ggtgtaaaaa gtactcaaaa attttactca agtgaaagta
       61 caagtactta gggaaaattt tactcaatta aaagtaaaag tatctggcta gaatcttact
      121 tgagtaaaag taaaaaagta ctccattaaa attgtacttg agtattaagg aagtaaaagt
      181 aaaagcaaga aagaaaacta gagattcttg tttaagcttt taatctcaaa aaacattaaa
      241 tgaaatgcat acaaggtttt atcctgcttt agaactgttt gtatttaatt atcaaactat
      301 aagacagaca atctaatgcc agtacacgct actcaaagtt gtaaaacctc agatttaact
      361 tcagtagaag ctgattctca aaattgttag tgtcaagcct agctcttttg gggctgaaaa
      421 gcaatcctgc agtgctgaaa agcctctcac aggcagccga tgcgggaaga ggtgtattag
      481 tcttgataga gaggctgcaa atagcaggaa acgtgagcag agactccctg gtgtctgaaa
      541 cacaggccag atgggccctc gacggtatcg ataagcttgc atgcctgcag gtcggagtac
      601 tgtcctccga gcggagtact gtcctccgag cggagtactg tcctccgagc ggagtactgt
      661 cctccgagcg gagtactgtc ctccgagcgg agactctaga gggtatataa tggatcccca
      721 ccatggcgcg gctgctgccc gcgctgctgc tgctgctgct gcccggtaac gtgaccggta
      781 acgggtccgg taacggttct ttgtcccgtt gcccccccgg tcagttccgc tgctcggagc
      841 cgcccggtgc ccacggggag tgttacccgc aggactggct gtgcgacgga caccccgact
      901 gcgacgacgg gcgggacgag tggggctgcg ggaccagcgc gatccccgcg gtgcccacgg
      961 acaacggcac agaggctccc actgcccctg ctcctggacg tgctctgcca gccaggaatc
     1021 acggccgcat gtggatgctg atcactgcag tgctcctgtg ctgcctggta gctgtgggtg
     1081 gtatcgctgc atgggggaag tccaaagcaa aaagcaggtc tgacatcttc agtcttgaaa
     1141 gcgcatccaa ggggggtggg gtgagcaagg gcgaggagga taacatggcc atcatcaagg
     1201 agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg
     1261 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
     1321 agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca
     1381 aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg
     1441 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg
     1501 actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc
     1561 cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga
     1621 tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg
     1681 gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc
     1741 ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca
     1801 tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt
     1861 acaagtaatc tagacagagt cgctgatcag cctcgactgt gccttctagt tgccagccat
     1921 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     1981 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     2041 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgcaa
     2101 gctgctagct agggataaca gggtaataga tctgcgaaga tacggccacg ggtgctcttg
     2161 atcctgtggc tgattttgga ctgtgctgct cgcagctgct gatgaatcac atacttcctc
     2221 cattttcttc cactgattga ctgttataat ttccctaatt tccaggtcaa ggtgctgtgc
     2281 attgtggtaa tagatgtgac atgacgtcac ttccaaagga ccaatgaaca tgtctgacca
     2341 atttcatata atgtgaaaac gattttcata ggcagaataa ataacattta aattaaactg
     2401 ggcatcagcg caattcaatt ggtttggtaa tagcaaggga aaatagaatg aagtgatctc
     2461 caaaaaataa gtactttttg actgtaaata aaattgtaag gagtaaaaag tacttttttt
     2521 tctaaaaaaa tgtaattaag taaaagtaaa agtattgatt tttaattgta ctcaagtaaa
     2581 gtaaaaatcc ccaaaaataa tacttaagta cagtaatcaa gtaaaattac tcaagtactt
     2641 tacacctctg gttcttgacc ccctaccttc agcaagccca gcagatccac tagggccgcc
     2701 accgcggtgg agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
     2761 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     2821 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     2881 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     2941 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     3001 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3061 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3121 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3181 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3241 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3301 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3361 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3421 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3481 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3541 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3601 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3661 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3721 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3781 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3841 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3901 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3961 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4021 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     4081 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4141 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4201 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4261 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4321 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     4381 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4441 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4501 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4561 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4621 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4681 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4741 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4801 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4861 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa
     4921 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     4981 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     5041 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     5101 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     5161 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     5221 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     5281 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     5341 cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac
     5401 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     5461 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     5521 acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattggg tacc
//
