LOCUS       sequence-407281-        8012 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vasa (closed conformation) expression for Schneider cell imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8012)
  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 8012)
  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: F16-pAc5.1 EGFP Vasa closed
FEATURES             Location/Qualifiers
     source          1..8012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..2517
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2537..2539,2540..2542,2543..3253)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2537..2539,2540..2542,2543..3253)
                     /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(2582..2603)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2843..2862)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3190..3211
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(3612..3629)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             5270..5311
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5321..5338
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5361..5378)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5411..5545
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5435..5454)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5489..5508
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6146..6734)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6226..6245)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6905..7696,7697..7765))
                     /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     7528..7547
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7766..7870)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7938..7956
                     /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 agagagagaa
     1561 aaagagggag agcagcttag accgcatgtg cttgtgtgtg aggcgtctct ctcttcgtct
     1621 ctgttgcgca aacgcataga ctgcactgaa aaaatcgatt acctattttt tatgaatgaa
     1681 tatttgcact attactattc aaaactatta agatagcaat cacattcaat agccaaatac
     1741 tataccacct gagcgatgca acgaaatgat caatttgagc aaaaatgctg catatttagg
     1801 acggcatcat tatagaaatg cttcttgctg tgtacttttc tctcgtctgg cagctgtttc
     1861 gccgttattg ttaaaaccgg cttaagttag gtgtgttttc tacgactagt gaatgcccta
     1921 ctagaagatg tgtgttgcac aaaatgtccc tggaataacc aatttgaagt gcagatagca
     1981 gtaaacgtaa gctaatatga atattattta actgtaatgt tttaatatcg ctggacatta
     2041 ctaataaacc cactataaac acatgtacat atgtatgttt tggcatacaa tgagtagttg
     2101 gggaaaaaat gtgtaaaagc accgtgacca tcacagcata aagataacca gctgaagtat
     2161 cgaatatgag taacccccaa attgaatcac atgccgcaac tgataggacc catggaagta
     2221 cactcttcat ggcgatatac aagacacaca caagcacgaa cacccagttg cggaggaaat
     2281 tctccgtaaa tgaaaaccca atcggcgaac aattcatacc catatatggt aaaagttttg
     2341 aacgcgactt gagagcggag agcattgcgg ctgataaggt tttagcgcta agcgggcttt
     2401 ataaaacggg ctgcgggacc agttttcata tcactaccgt ttgagttctt gtgctgtgtg
     2461 gatactcctc ccgacacaaa gccgctccat cagccagcag tcgtctaatc cagagacccc
     2521 ggatcggggt accatcatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     2581 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     2641 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     2701 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     2761 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     2821 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     2881 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     2941 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     3001 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     3061 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     3121 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     3181 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     3241 cgagctgtac aagccaagct tgaattctgc agatatgtct gacgactggg atgatgagcc
     3301 cattgttgat actcgcggcg cccgcggtgg agattggagc gatgatgagg acacggccaa
     3361 gagcttcagc ggcgaagctg aaggcgatgg tgttggaggg agcggtggtg aaggcggcgg
     3421 ctaccaagga ggaaatcgag atgtgttcgg aaggatcggc ggaggcagag gaggaggagc
     3481 tggaggttat cgaggaggaa atcgagatgg agggggcttc cacggtggac gtcgcgaggg
     3541 agaaagggac ttccgcggtg gagaaggcgg cttccgcggt ggacaaggcg gctcccgcgg
     3601 tggacaaggc ggctcccgcg ggggacaagg cggcttccgt ggtggagaag gcggcttccg
     3661 cggtcggctg tacgaaaacg aggatggtga tgaacgtcgt ggaagactag atcgggagga
     3721 acgcggcggt gaacgtggtg aaagagggga cggtggtttt gcgaggcgcc gtcgaaatga
     3781 agatgatata aataataata ataatatcgc cgaagatgtc gaaaggaagc gagagtttta
     3841 tattcctccc gagccgtcca acgatgcaat tgagatattc agcagtggca tcgcttccgg
     3901 gattcatttt tcgaaataca acaacatacc ggttaaggtg accggcagcg atgttccaca
     3961 acctatacaa cattttacga gcgctgattt gagagatatt attatagaca atgtaaacaa
     4021 atctggatac aaaataccga caccaataca aaagtgttcc ataccagtaa tatcctctgg
     4081 tcgagaccta atggcgtgtg cccagacggg ttctggaaaa acggcagctt ttttgttgcc
     4141 cattttgagc aagttgttgg aggatcccca tgaactggag cttggaagac cccaggtagt
     4201 gattgtatcc ccaacaaggg agctggcaat tcagatcttt aacgaggcga ggaagtttgc
     4261 tttcgagtca tatctaaaga tcggtattgt ttacggaggc acctcgttca gacatcaaaa
     4321 cgagtgcatt accagaggct gccatgtagt gatcgccact ccgggacgac ttctggattt
     4381 cgtggatcgg acctttatca cgtttgaaga cactcgattc gttgtgctgg atcaggccga
     4441 tcgcatgcta gatatgggtt tctcagaaga catgcgaaga atcatgacgc atgtaactat
     4501 gcgtccagaa catcagacat tgatgttttc cgccacgttt ccagaagaaa tccaaagaat
     4561 ggccggcgaa ttccttaaaa attacgtttt cgtcgccatt ggcattgtag gcggagcttg
     4621 ctctgatgtg aagcagacca tatacgaagt taataagtac gccaagcgat ccaagctaat
     4681 agaaatcctt tccgagcaag cagatggcac cattgtgttt gtggagacaa agcgtggcgc
     4741 cgacttttta gcttccttcc tgtcagaaaa ggagtttccg acgacctcca ttcatggcga
     4801 tcgtctccag agtcaacgcg agcaggcctt gcgtgatttc aagaacggct ctatgaaggt
     4861 tctcatagcc acttcagtgg cttctcgtgg attagacatt aaaaacatca agcatgtgat
     4921 caactatgac atgcccagca agatcgatga ctacgtacat cgcattggac gtacaggtcg
     4981 tgtaggcaat aatggacgag ccacaagctt ctttgaccct gaaaaggatc gagctattgc
     5041 tgcggacttg gtaaaaatct tggagggatc tggccagact gttccggact ttctacgcac
     5101 ctgtggtgcc ggcggtgatg ggggttactc caatcaaaat ttcggtgggg tggatgtgcg
     5161 tggcagggga aattacgtcg gcgatgccac caatgtcgag gaagaagagc aatgggattg
     5221 aatccagcac agtggcggcc gctcgagtct agagggcccg cggttcgaag gtaagcctat
     5281 ccctaaccct ctcctcggtc tcgattctac gcgtaccggt catcatcacc atcaccattg
     5341 agtttaaacc cgctgatcag cctcgactgt gccttctaag gcctgagctc gctgatcagc
     5401 ctcgatcgag gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     5461 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     5521 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     5581 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc
     5641 tgattatgat cagtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt
     5701 gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg
     5761 taggacaggt gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg
     5821 cgacgatgat cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct
     5881 tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg
     5941 cggccgacgc gctgggctac gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc
     6001 ccattatgat tcttctcgct tccggcggca tcgggatgcc cgcgttgcag gccatgctgt
     6061 ccaggcaggt agatgacgac catcagggac agcttcaagg ccagcaaaag gccaggaacc
     6121 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6181 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6241 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6301 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6361 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6421 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6481 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6541 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     6601 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6661 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6721 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6781 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6841 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6901 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6961 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7021 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     7081 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     7141 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     7201 gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     7261 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7321 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7381 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7441 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7501 gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag
     7561 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7621 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7681 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7741 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7801 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7861 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     7921 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa gaattaattc
     7981 tatattctaa aaacacaaat gatacttcta aa
//
