LOCUS       sequence-407099-        8010 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vasa expression in Schneider cells for imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8010)
  AUTHORS   Salgania HK, Metz J, Jeske M
  TITLE     ReLo is a simple and rapid colocalization assay to identify and 
            characterize direct protein-protein interactions.
  JOURNAL   Nat Commun. 2024 Apr 3;15(1):2875. doi: 10.1038/s41467-024-47233-4.
  PUBMED    38570497
REFERENCE   2  (bases 1 to 8010)
  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: "Nat 
            Commun."; date: "2024-04-3"; volume: "15(1):2875. doi"; pages: " 
            10.1038/s41467-024-47233-4"
COMMENT     Sequence Label: pAc5.1-EGFP-Vasa open (K295N) (RR228)
FEATURES             Location/Qualifiers
     source          1..8010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..2515
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2535..2537,2538..2540,2541..3251)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2535..2537,2538..2540,2541..3251)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2580..2601)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2841..2860)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3188..3209
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(3610..3627)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             5268..5309
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5319..5336
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5359..5376)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5409..5543
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5433..5452)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5487..5506
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6144..6732)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6224..6243)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6903..7694,7695..7763))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7526..7545
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7764..7868)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7936..7954
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aaaaaatcat gaatggcatc aactctgaat caaatctttg cagatgcacc tacttctcat
       61 ttccactgtc acatcatttt tccagatctc gctgcctgtt atgtggccca caaaccaaga
      121 cacgttttat ggccattaaa gctggctgat cgtcgccaaa caccaaatac ataatgaata
      181 tgtacacatt cgagaaagaa gcgatcaaag aagcgtcttc gggcggagta ggagaatgcg
      241 gaggagaagg agaacgagct gatctagtat ctctccacaa tccaatgcca actgaccaac
      301 tggccatatt cggagcaatt tgaagccaat ttccatcgcc tggcgatcgc tccattcttg
      361 gctatatgtt tttcaccgtt acccggggcc attttcaaag actcgtcggc aagataagat
      421 tgtgtcactc gctgtctctc ttcatttgtc gaagaatgct gaggaatttc gcgatgacgt
      481 cggcgagtat tttgaagaat gagaataatt tgtatttata cgaaaatcag ttagtggaat
      541 tttctacaaa aacatgttat ctatagataa ttttgttgca aaatatgttg actatgacaa
      601 agattgtatg tatatacctt taatgtattc tcattttctt atgtatttat aatggcaatg
      661 atgatactga tgatatttta agatgatgcc agaccaaaag gcttgaattt ctgcgtcttt
      721 tgccgaacgc agtgcatgtg caattgttgt tttttggaat attcaatttt cggactgtcc
      781 gctttgattt cagtttcttg gcttattcaa aaagcaaagt aaagccaaaa aagcgagatg
      841 gcaataccaa atgcggcaaa acggtagtgg aaggaaaggg gtgcggggca gcggaaggaa
      901 gggtggggcg gggcgtggcg gggtctgtgg ctgggcgcga cgtcaccgac gttggagcca
      961 ctcctttgac catgtgtgcg tgtgtgtatt attcgtgtct cgccactcgc cggttgtttt
     1021 tttcttttta tgctgcgctc tctctagcgc catctcgctt acgcatgctc aacgcaccgc
     1081 atgttgccgt ttccttttat gcgtcatttt ggctcgaaat aggcaattat ttaaacaaag
     1141 attagtcaac gaaaacgcta aaataaataa gtctacaata tggttactta ttgccatgtg
     1201 tgtgcagcca acgatagcaa caaaagcaac aacacaggtg gctttccctc tttcactttt
     1261 tgtttgcaag ccgcgtgcga gcaagacggc acgaccggca aacgcaatta cgctgacaaa
     1321 gagcagacga agttttggcg aaaaacatca aggcgcctga tacgaatgca tttgcaataa
     1381 caattgcgat atttaatatt gtttatgaag ctgtttgact tcaaaacaca caaaaaaaaa
     1441 aataaaacaa attatttgaa agagaattag gaatcggacg cttatcgtta gggtaacaac
     1501 aagaaatgct tactgagtca cagcctctgg aaaactgccg caagccagag agagagaaaa
     1561 agagggagag cagcttagac cgcatgtgct tgtgtgtgag gcgtctctct cttcgtctct
     1621 gttgcgcaaa cgcatagact gcactgaaaa aatcgattac ctatttttta tgaatgaata
     1681 tttgcactat tactattcaa aactattaag atagcaatca cattcaatag ccaaatacta
     1741 taccacctga gcgatgcaac gaaatgatca atttgagcaa aaatgctgca tatttaggac
     1801 ggcatcatta tagaaatgct tcttgctgtg tacttttctc tcgtctggca gctgtttcgc
     1861 cgttattgtt aaaaccggct taagttaggt gtgttttcta cgactagtga atgccctact
     1921 agaagatgtg tgttgcacaa aatgtccctg gaataaccaa tttgaagtgc agatagcagt
     1981 aaacgtaagc taatatgaat attatttaac tgtaatgttt taatatcgct ggacattact
     2041 aataaaccca ctataaacac atgtacatat gtatgttttg gcatacaatg agtagttggg
     2101 gaaaaaatgt gtaaaagcac cgtgaccatc acagcataaa gataaccagc tgaagtatcg
     2161 aatatgagta acccccaaat tgaatcacat gccgcaactg ataggaccca tggaagtaca
     2221 ctcttcatgg cgatatacaa gacacacaca agcacgaaca cccagttgcg gaggaaattc
     2281 tccgtaaatg aaaacccaat cggcgaacaa ttcataccca tatatggtaa aagttttgaa
     2341 cgcgacttga gagcggagag cattgcggct gataaggttt tagcgctaag cgggctttat
     2401 aaaacgggct gcgggaccag ttttcatatc actaccgttt gagttcttgt gctgtgtgga
     2461 tactcctccc gacacaaagc cgctccatca gccagcagtc gtctaatcca gagaccccgg
     2521 atcggggtac catcatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     2581 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     2641 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     2701 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     2761 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     2821 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     2881 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     2941 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     3001 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     3061 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     3121 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     3181 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     3241 agctgtacaa gccaagcttg aattctgcag atatgtctga cgactgggat gatgagccca
     3301 ttgttgatac tcgcggcgcc cgcggtggag attggagcga tgatgaggac acggccaaga
     3361 gcttcagcgg cgaagctgaa ggcgatggtg ttggagggag cggtggtgaa ggcggcggct
     3421 accaaggagg aaatcgagat gtgttcggaa ggatcggcgg aggcagagga ggaggagctg
     3481 gaggttatcg aggaggaaat cgagatggag ggggcttcca cggtggacgt cgcgagggag
     3541 aaagggactt ccgcggtgga gaaggcggct tccgcggtgg acaaggcggc tcccgcggtg
     3601 gacaaggcgg ctcccgcggg ggacaaggcg gcttccgtgg tggagaaggc ggcttccgcg
     3661 gtcggctgta cgaaaacgag gatggtgatg aacgtcgtgg aagactagat cgggaggaac
     3721 gcggcggtga acgtggtgaa agaggggacg gtggttttgc gaggcgccgt cgaaatgaag
     3781 atgatataaa taataataat aatatcgccg aagatgtcga aaggaagcga gagttttata
     3841 ttcctcccga gccgtccaac gatgcaattg agatattcag cagtggcatc gcttccggga
     3901 ttcatttttc gaaatacaac aacataccgg ttaaggtgac cggcagcgat gttccacaac
     3961 ctatacaaca ttttacgagc gctgatttga gagatattat tatagacaat gtaaacaaat
     4021 ctggatacaa aataccgaca ccaatacaaa agtgttccat accagtaata tcctctggtc
     4081 gagacctaat ggcgtgtgcc cagacgggtt ctggaaacac ggcagctttt ttgttgccca
     4141 ttttgagcaa gttgttggag gatccccatg aactggagct tggaagaccc caggtagtga
     4201 ttgtatcccc aacaagggag ctggcaattc agatctttaa cgaggcgagg aagtttgctt
     4261 tcgagtcata tctaaagatc ggtattgttt acggaggcac ctcgttcaga catcaaaacg
     4321 agtgcattac cagaggctgc catgtagtga tcgccactcc gggacgactt ctggatttcg
     4381 tggatcggac ctttatcacg tttgaagaca ctcgattcgt tgtgctggat gaggccgatc
     4441 gcatgctaga tatgggtttc tcagaagaca tgcgaagaat catgacgcat gtaactatgc
     4501 gtccagaaca tcagacattg atgttttccg ccacgtttcc agaagaaatc caaagaatgg
     4561 ccggcgaatt ccttaaaaat tacgttttcg tcgccattgg cattgtaggc ggagcttgct
     4621 ctgatgtgaa gcagaccata tacgaagtta ataagtacgc caagcgatcc aagctaatag
     4681 aaatcctttc cgagcaagca gatggcacca ttgtgtttgt ggagacaaag cgtggcgccg
     4741 actttttagc ttccttcctg tcagaaaagg agtttccgac gacctccatt catggcgatc
     4801 gtctccagag tcaacgcgag caggccttgc gtgatttcaa gaacggctct atgaaggttc
     4861 tcatagccac ttcagtggct tctcgtggat tagacattaa aaacatcaag catgtgatca
     4921 actatgacat gcccagcaag atcgatgact acgtacatcg cattggacgt acaggtcgtg
     4981 taggcaataa tggacgagcc acaagcttct ttgaccctga aaaggatcga gctattgctg
     5041 cggacttggt aaaaatcttg gagggatctg gccagactgt tccggacttt ctacgcacct
     5101 gtggtgccgg cggtgatggg ggttactcca atcaaaattt cggtggggtg gatgtgcgtg
     5161 gcaggggaaa ttacgtcggc gatgccacca atgtcgagga agaagagcaa tgggattgaa
     5221 tccagcacag tggcggccgc tcgagtctag agggcccgcg gttcgaaggt aagcctatcc
     5281 ctaaccctct cctcggtctc gattctacgc gtaccggtca tcatcaccat caccattgag
     5341 tttaaacccg ctgatcagcc tcgactgtgc cttctaaggc ctgagctcgc tgatcagcct
     5401 cgatcgagga tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     5461 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     5521 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     5581 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg
     5641 attatgatca gtcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg
     5701 gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta
     5761 ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg
     5821 acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc acgccctcgc tcaagccttc
     5881 gtcactggtc ccgccaccaa acgtttcggc gagaagcagg ccattatcgc cggcatggcg
     5941 gccgacgcgc tgggctacgt cttgctggcg ttcgcgacgc gaggctggat ggccttcccc
     6001 attatgattc ttctcgcttc cggcggcatc gggatgcccg cgttgcaggc catgctgtcc
     6061 aggcaggtag atgacgacca tcagggacag cttcaaggcc agcaaaaggc caggaaccgt
     6121 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6181 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6241 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6301 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6361 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6421 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6481 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6541 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     6601 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6661 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6721 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6781 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6841 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6901 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6961 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7021 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7081 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7141 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7201 aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7261 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7321 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7381 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7441 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7501 tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg
     7561 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7621 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7681 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7741 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7801 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7861 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     7921 acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcaaga attaattcta
     7981 tattctaaaa acacaaatga tacttctaaa
//
