LOCUS       sequence-407278-        6857 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vasa 200-642 expression for Schneider cell imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6857)
  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 6857)
  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: F18-pAc5.1 EGFP Vasa 200-462
FEATURES             Location/Qualifiers
     source          1..6857
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..2565
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2585..2587,2588..2590,2591..3301)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2585..2587,2588..2590,2591..3301)
                     /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(2630..2651)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2891..2910)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3238..3259
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4163..4204
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4214..4231
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4254..4271)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4304..4438
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4328..4347)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4382..4401
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(5039..5627)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5119..5138)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5798..6589,6590..6658))
                     /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     6421..6440
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6659..6763)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6831..6849
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cttcaagaat taattctata ttctaaaaac acaaatgata cttctaaaaa aaaatcatga
       61 atggcatcaa ctctgaatca aatctttgca gatgcaccta cttctcattt ccactgtcac
      121 atcatttttc cagatctcgc tgcctgttat gtggcccaca aaccaagaca cgttttatgg
      181 ccattaaagc tggctgatcg tcgccaaaca ccaaatacat aatgaatatg tacacattcg
      241 agaaagaagc gatcaaagaa gcgtcttcgg gcggagtagg agaatgcgga ggagaaggag
      301 aacgagctga tctagtatct ctccacaatc caatgccaac tgaccaactg gccatattcg
      361 gagcaatttg aagccaattt ccatcgcctg gcgatcgctc cattcttggc tatatgtttt
      421 tcaccgttac ccggggccat tttcaaagac tcgtcggcaa gataagattg tgtcactcgc
      481 tgtctctctt catttgtcga agaatgctga ggaatttcgc gatgacgtcg gcgagtattt
      541 tgaagaatga gaataatttg tatttatacg aaaatcagtt agtggaattt tctacaaaaa
      601 catgttatct atagataatt ttgttgcaaa atatgttgac tatgacaaag attgtatgta
      661 tataccttta atgtattctc attttcttat gtatttataa tggcaatgat gatactgatg
      721 atattttaag atgatgccag accaaaaggc ttgaatttct gcgtcttttg ccgaacgcag
      781 tgcatgtgca attgttgttt tttggaatat tcaattttcg gactgtccgc tttgatttca
      841 gtttcttggc ttattcaaaa agcaaagtaa agccaaaaaa gcgagatggc aataccaaat
      901 gcggcaaaac ggtagtggaa ggaaaggggt gcggggcagc ggaaggaagg gtggggcggg
      961 gcgtggcggg gtctgtggct gggcgcgacg tcaccgacgt tggagccact cctttgacca
     1021 tgtgtgcgtg tgtgtattat tcgtgtctcg ccactcgccg gttgtttttt tctttttatg
     1081 ctgcgctctc tctagcgcca tctcgcttac gcatgctcaa cgcaccgcat gttgccgttt
     1141 ccttttatgc gtcattttgg ctcgaaatag gcaattattt aaacaaagat tagtcaacga
     1201 aaacgctaaa ataaataagt ctacaatatg gttacttatt gccatgtgtg tgcagccaac
     1261 gatagcaaca aaagcaacaa cacaggtggc tttccctctt tcactttttg tttgcaagcc
     1321 gcgtgcgagc aagacggcac gaccggcaaa cgcaattacg ctgacaaaga gcagacgaag
     1381 ttttggcgaa aaacatcaag gcgcctgata cgaatgcatt tgcaataaca attgcgatat
     1441 ttaatattgt ttatgaagct gtttgacttc aaaacacaca aaaaaaaaaa taaaacaaat
     1501 tatttgaaag agaattagga atcggacgct tatcgttagg gtaacaacaa gaaatgctta
     1561 ctgagtcaca gcctctggaa aactgccgca agccagagag agagagaaaa agagggagag
     1621 cagcttagac cgcatgtgct tgtgtgtgag gcgtctctct cttcgtctct gttgcgcaaa
     1681 cgcatagact gcactgaaaa aatcgattac ctatttttta tgaatgaata tttgcactat
     1741 tactattcaa aactattaag atagcaatca cattcaatag ccaaatacta taccacctga
     1801 gcgatgcaac gaaatgatca atttgagcaa aaatgctgca tatttaggac ggcatcatta
     1861 tagaaatgct tcttgctgtg tacttttctc tcgtctggca gctgtttcgc cgttattgtt
     1921 aaaaccggct taagttaggt gtgttttcta cgactagtga atgccctact agaagatgtg
     1981 tgttgcacaa aatgtccctg gaataaccaa tttgaagtgc agatagcagt aaacgtaagc
     2041 taatatgaat attatttaac tgtaatgttt taatatcgct ggacattact aataaaccca
     2101 ctataaacac atgtacatat gtatgttttg gcatacaatg agtagttggg gaaaaaatgt
     2161 gtaaaagcac cgtgaccatc acagcataaa gataaccagc tgaagtatcg aatatgagta
     2221 acccccaaat tgaatcacat gccgcaactg ataggaccca tggaagtaca ctcttcatgg
     2281 cgatatacaa gacacacaca agcacgaaca cccagttgcg gaggaaattc tccgtaaatg
     2341 aaaacccaat cggcgaacaa ttcataccca tatatggtaa aagttttgaa cgcgacttga
     2401 gagcggagag cattgcggct gataaggttt tagcgctaag cgggctttat aaaacgggct
     2461 gcgggaccag ttttcatatc actaccgttt gagttcttgt gctgtgtgga tactcctccc
     2521 gacacaaagc cgctccatca gccagcagtc gtctaatcca gagaccccgg atcggggtac
     2581 catcatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     2641 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     2701 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     2761 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     2821 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     2881 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     2941 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     3001 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     3061 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     3121 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     3181 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     3241 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     3301 gccaagcttg aattctgcag atgagtttta tattcctccc gagccgtcca acgatgcaat
     3361 tgagatattc agcagtggca tcgcttccgg gattcatttt tcgaaataca acaacatacc
     3421 ggttaaggtg accggcagcg atgttccaca acctatacaa cattttacga gcgctgattt
     3481 gagagatatt attatagaca atgtaaacaa atctggatac aaaataccga caccaataca
     3541 aaagtgttcc ataccagtaa tatcctctgg tcgagaccta atggcgtgtg cccagacggg
     3601 ttctggaaaa acggcagctt ttttgttgcc cattttgagc aagttgttgg aggatcccca
     3661 tgaactggag cttggaagac cccaggtagt gattgtatcc ccaacaaggg agctggcaat
     3721 tcagatcttt aacgaggcga ggaagtttgc tttcgagtca tatctaaaga tcggtattgt
     3781 ttacggaggc acctcgttca gacatcaaaa cgagtgcatt accagaggct gccatgtagt
     3841 gatcgccact ccgggacgac ttctggattt cgtggatcgg acctttatca cgtttgaaga
     3901 cactcgattc gttgtgctgg atgaggccga tcgcatgcta gatatgggtt tctcagaaga
     3961 catgcgaaga atcatgacgc atgtaactat gcgtccagaa catcagacat tgatgttttc
     4021 cgccacgttt ccagaagaaa tccaaagaat ggccggcgaa ttccttaaaa attacgtttt
     4081 cgtcgccatt ggcattgtag gcggagcttg ctagatccag cacagtggcg gccgctcgag
     4141 tctagagggc ccgcggttcg aaggtaagcc tatccctaac cctctcctcg gtctcgattc
     4201 tacgcgtacc ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac
     4261 tgtgccttct aaggcctgag ctcgctgatc agcctcgatc gaggatccag acatgataag
     4321 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     4381 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     4441 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     4501 aagcaagtaa aacctctaca aatgtggtat ggctgattat gatcagtcga ccgatgccct
     4561 tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg
     4621 cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg
     4681 tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg
     4741 tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt
     4801 tcggcgagaa gcaggccatt atcgccggca tggcggccga cgcgctgggc tacgtcttgc
     4861 tggcgttcgc gacgcgaggc tggatggcct tccccattat gattcttctc gcttccggcg
     4921 gcatcgggat gcccgcgttg caggccatgc tgtccaggca ggtagatgac gaccatcagg
     4981 gacagcttca aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5041 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5101 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5161 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5221 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5281 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5341 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5401 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5461 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5521 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5581 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5641 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5701 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5761 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5821 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5881 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5941 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6001 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6061 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca
     6121 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6181 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6241 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6301 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6361 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaacacggg
     6421 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6481 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6541 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6601 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6661 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6721 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6781 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     6841 tcacgaggcc ctttcgt
//
