LOCUS       sequence-267227-        5241 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Venus knock-in cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5241)
  AUTHORS   Kondo S, Takahashi T, Yamagata N, Imanishi Y, Katow H, Hiramatsu S, 
            Lynn K, Abe A, Kumaraswamy A, Tanimoto H
  TITLE     Neurochemical Organization of the Drosophila Brain Visualized by 
            Endogenously Tagged Neurotransmitter Receptors.
  JOURNAL   Cell Rep. 2020 Jan 7;30(1):284-297.e5. doi: 
            10.1016/j.celrep.2019.12.018.
  PUBMED    31914394
REFERENCE   2  (bases 1 to 5241)
  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: 
            "10.1016/j.celrep.2019.12"; journalName: "Cell Rep"; date: 
            "2020-01-7- 7"; volume: "30"; issue: "1"; pages: "284-297.e5"
COMMENT     Sequence Label: pPVxRF3
FEATURES             Location/Qualifiers
     source          1..5241
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(77..79,80..82,83..796)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(122..143)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(383..402)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     730..751
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(805..838)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             1376..2053
                     /codon_start=1
                     /product="noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2008)"
                     /label=DsRed-Express2
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     complement(1556..1576)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1774..1793
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1968..1991
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2169..2250
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2206..2225)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2260..2279
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2294..2327)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2334..2367
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     primer_bind     complement(2401..2421)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2401..2419)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2440..2456)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2440..2456)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2453..2475)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2464..2480
                     /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(2488..2494,2495..2512,2513..2518))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2533..2554
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2671..2688)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2842..3430)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2922..2941)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3601..4392,4393..4461))
                     /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     4224..4243
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4462..4566)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4593..5048
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4680..4699)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4890..4911
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5174..5196
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5188..5205
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5189..5205
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5215..5234
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5215..5233
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggtacccgtc tcacctacgc ccccaactga gagaactcaa aggttacccc agttggggca
       61 ctacggcgcg cccaaaatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
      121 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
      181 gggcgatgcc acctacggca agctgaccct gaagctcatc tgcaccaccg gcaagctgcc
      241 cgtgccctgg cccaccctcg tgaccaccct gggctacggc ctgcagtgct tcgcccgcta
      301 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
      361 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
      421 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
      481 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc
      541 cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg
      601 cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
      661 gctgcccgac aaccactacc tgagctacca gtccgccctg agcaaagacc ccaacgagaa
      721 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
      781 cgagctgtac aagtaagcgg ccgcataact tcgtatagca tacattatac gaagttatat
      841 tcgaatcgga aatcaatcga attctcgatc aatgttttgc gaaaaaaaat gaaacgagaa
      901 caaatgaatt cagttgtaaa agatttgttc tcttaacaac gtaatcaata aagtcggatc
      961 caattttgtt cacaagtcta aacaattatg gtttttgttg gggtttctta taattaataa
     1021 gagagatatc aatcatttta cgaggattat gagactgcgt tttgtagtcc aagaaagctg
     1081 attaatgttt agagtgttgg tataaatgtt aataggctca ttaatgtagg gcatgcgacg
     1141 tcaggcctaa gcttgagctc gcccggggat ctaattcaat tagagactaa ttcaattaga
     1201 gctaattcaa ttaggatcag cttatcgatt tcgaaccctc gaccgccgga gtataaatag
     1261 aggcgcttcg tctacggagc gacaattcaa ttcaaacaag caaagtgaac acgtcgctaa
     1321 gcgaaagcta agcaaataaa caagcgcagc tgaacaagct aaacaatcgg gcaaaatgga
     1381 tagcactgag aacgtcatca agcccttcat gcgcttcaag gtgcacatgg agggctccgt
     1441 gaacggccac gagttcgaga tcgagggcga gggcgagggc aagccctacg agggcaccca
     1501 gaccgccaag ctgcaggtga ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc
     1561 cccccagttc cagtacggct ccaaggtgta cgtgaagcac cccgccgaca tccccgacta
     1621 caagaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     1681 cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcaccttca tctaccacgt
     1741 gaagttcatc ggcgtgaact tcccctccga cggccccgta atgcagaaga agactctggg
     1801 ctgggagccc tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca
     1861 caaggcgctg aagctgaagg gcggcggcca ctacctggtg gagttcaagt caatctacat
     1921 ggccaagaag cccgtgaagc tgcccggcta ctactacgtg gactccaagc tggacatcac
     1981 ctcccacaac gaggactaca ccgtggtgga gcagtacgag cgcgccgagg cccgccacca
     2041 cctgttccag tagccatggc tagatcataa tcagccatac cacatttgta gaggttttac
     2101 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     2161 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     2221 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     2281 atgtatcaag cttataactt cgtatagcat acattatacg aagttatatg catgaagttc
     2341 ctattcttca aatagtatag gaacttccga gacgccgcgg tggagctcca gcttttgttc
     2401 cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     2461 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     2521 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     2581 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     2641 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2701 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2761 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2821 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2881 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2941 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3001 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3061 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3121 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3181 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3241 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     3301 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3361 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3421 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3481 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3541 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3601 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3661 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3721 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3781 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3841 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3901 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3961 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4021 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4081 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4141 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4201 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4261 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4321 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4381 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4441 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4501 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4561 tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc
     4621 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4681 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4741 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4801 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4861 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4921 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     4981 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     5041 tacaatttcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     5101 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     5161 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac
     5221 gactcactat agggcgaatt g
//
