LOCUS       sequence-275173-        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"
     primer_bind     17..39
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     31..48
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     32..48
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     58..77
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        58..76
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(161..163,164..166,167..880)
                     /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(206..227)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(467..486)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     814..835
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(889..922)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             1460..2137
                     /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(1640..1660)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1858..1877
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2052..2075
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2253..2334
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2290..2309)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2344..2363
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2378..2411)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2418..2451
                     /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(2485..2505)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2485..2503)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2524..2540)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2524..2540)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2537..2559)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2548..2564
                     /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(2572..2578,2579..2596,2597..2602))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2617..2638
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2755..2772)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2926..3514)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3006..3025)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3685..4476,4477..4545))
                     /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     4308..4327
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4546..4650)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4676..5131)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4813..4834)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5025..5044
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa
       61 tacgactcac tatagggcga attgggtacc cgtctcacct acgcccccaa ctgagagaac
      121 tcaaaggtta ccccagttgg ggcactacgg cgcgcccaaa atggtgagca agggcgagga
      181 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
      241 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct
      301 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta
      361 cggcctgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
      421 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
      481 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
      541 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
      601 cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa
      661 gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac
      721 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccgc
      781 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
      841 cgccgggatc actctcggca tggacgagct gtacaagtaa gcggccgcat aacttcgtat
      901 agcatacatt atacgaagtt atattcgaat cggaaatcaa tcgaattctc gatcaatgtt
      961 ttgcgaaaaa aaatgaaacg agaacaaatg aattcagttg taaaagattt gttctcttaa
     1021 caacgtaatc aataaagtcg gatccaattt tgttcacaag tctaaacaat tatggttttt
     1081 gttggggttt cttataatta ataagagaga tatcaatcat tttacgagga ttatgagact
     1141 gcgttttgta gtccaagaaa gctgattaat gtttagagtg ttggtataaa tgttaatagg
     1201 ctcattaatg tagggcatgc gacgtcaggc ctaagcttga gctcgcccgg ggatctaatt
     1261 caattagaga ctaattcaat tagagctaat tcaattagga tcagcttatc gatttcgaac
     1321 cctcgaccgc cggagtataa atagaggcgc ttcgtctacg gagcgacaat tcaattcaaa
     1381 caagcaaagt gaacacgtcg ctaagcgaaa gctaagcaaa taaacaagcg cagctgaaca
     1441 agctaaacaa tcgggcaaaa tggatagcac tgagaacgtc atcaagccct tcatgcgctt
     1501 caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga
     1561 gggcaagccc tacgagggca cccagaccgc caagctgcag gtgaccaagg gcggccccct
     1621 gcccttcgcc tgggacatcc tgtcccccca gttccagtac ggctccaagg tgtacgtgaa
     1681 gcaccccgcc gacatccccg actacaagaa gctgtccttc cccgagggct tcaagtggga
     1741 gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca
     1801 ggacggcacc ttcatctacc acgtgaagtt catcggcgtg aacttcccct ccgacggccc
     1861 cgtaatgcag aagaagactc tgggctggga gccctccacc gagcgcctgt acccccgcga
     1921 cggcgtgctg aagggcgaga tccacaaggc gctgaagctg aagggcggcg gccactacct
     1981 ggtggagttc aagtcaatct acatggccaa gaagcccgtg aagctgcccg gctactacta
     2041 cgtggactcc aagctggaca tcacctccca caacgaggac tacaccgtgg tggagcagta
     2101 cgagcgcgcc gaggcccgcc accacctgtt ccagtagcca tggctagatc ataatcagcc
     2161 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     2221 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     2281 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     2341 gttgtggttt gtccaaactc atcaatgtat caagcttata acttcgtata gcatacatta
     2401 tacgaagtta tatgcatgaa gttcctattc ttcaaatagt ataggaactt ccgagacgcc
     2461 gcggtggagc tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     2521 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     2581 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     2641 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     2701 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     2761 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2821 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2881 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2941 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3001 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3061 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3121 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3181 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3241 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3301 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3361 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3421 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3481 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3541 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3601 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3661 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3721 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3781 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3841 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3901 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3961 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4021 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4081 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4141 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4201 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4261 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4321 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4381 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4441 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4501 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4561 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4621 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
     4681 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     4741 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     4801 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     4861 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     4921 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     4981 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     5041 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     5101 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt
     5161 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     5221 gctgcaaggc gattaagttg g
//
