LOCUS       sequence-407098-        6665 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vasa 463-661 expression in Schneider cells for imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6665)
  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 6665)
  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 463-661 MUT (F504E) (HK196)
FEATURES             Location/Qualifiers
     source          1..6665
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        9..2518
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2538..2540,2541..2543,2544..3254)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2538..2540,2541..2543,2544..3254)
                     /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(2583..2604)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2844..2863)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3191..3212
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3924..3965
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3975..3992
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4015..4032)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4065..4199
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4089..4108)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4143..4162
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4800..5388)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4880..4899)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5559..6350,6351..6419))
                     /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     6182..6201
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6420..6524)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6592..6610
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aaaaaaatca tgaatggcat caactctgaa tcaaatcttt gcagatgcac ctacttctca
       61 tttccactgt cacatcattt ttccagatct cgctgcctgt tatgtggccc acaaaccaag
      121 acacgtttta tggccattaa agctggctga tcgtcgccaa acaccaaata cataatgaat
      181 atgtacacat tcgagaaaga agcgatcaaa gaagcgtctt cgggcggagt aggagaatgc
      241 ggaggagaag gagaacgagc tgatctagta tctctccaca atccaatgcc aactgaccaa
      301 ctggccatat tcggagcaat ttgaagccaa tttccatcgc ctggcgatcg ctccattctt
      361 ggctatatgt ttttcaccgt tacccggggc cattttcaaa gactcgtcgg caagataaga
      421 ttgtgtcact cgctgtctct cttcatttgt cgaagaatgc tgaggaattt cgcgatgacg
      481 tcggcgagta ttttgaagaa tgagaataat ttgtatttat acgaaaatca gttagtggaa
      541 ttttctacaa aaacatgtta tctatagata attttgttgc aaaatatgtt gactatgaca
      601 aagattgtat gtatatacct ttaatgtatt ctcattttct tatgtattta taatggcaat
      661 gatgatactg atgatatttt aagatgatgc cagaccaaaa ggcttgaatt tctgcgtctt
      721 ttgccgaacg cagtgcatgt gcaattgttg ttttttggaa tattcaattt tcggactgtc
      781 cgctttgatt tcagtttctt ggcttattca aaaagcaaag taaagccaaa aaagcgagat
      841 ggcaatacca aatgcggcaa aacggtagtg gaaggaaagg ggtgcggggc agcggaagga
      901 agggtggggc ggggcgtggc ggggtctgtg gctgggcgcg acgtcaccga cgttggagcc
      961 actcctttga ccatgtgtgc gtgtgtgtat tattcgtgtc tcgccactcg ccggttgttt
     1021 ttttcttttt atgctgcgct ctctctagcg ccatctcgct tacgcatgct caacgcaccg
     1081 catgttgccg tttcctttta tgcgtcattt tggctcgaaa taggcaatta tttaaacaaa
     1141 gattagtcaa cgaaaacgct aaaataaata agtctacaat atggttactt attgccatgt
     1201 gtgtgcagcc aacgatagca acaaaagcaa caacacaggt ggctttccct ctttcacttt
     1261 ttgtttgcaa gccgcgtgcg agcaagacgg cacgaccggc aaacgcaatt acgctgacaa
     1321 agagcagacg aagttttggc gaaaaacatc aaggcgcctg atacgaatgc atttgcaata
     1381 acaattgcga tatttaatat tgtttatgaa gctgtttgac ttcaaaacac acaaaaaaaa
     1441 aaataaaaca aattatttga aagagaatta ggaatcggac gcttatcgtt agggtaacaa
     1501 caagaaatgc ttactgagtc acagcctctg gaaaactgcc gcaagccaga gagagagaga
     1561 aaaagaggga gagcagctta gaccgcatgt gcttgtgtgt gaggcgtctc tctcttcgtc
     1621 tctgttgcgc aaacgcatag actgcactga aaaaatcgat tacctatttt ttatgaatga
     1681 atatttgcac tattactatt caaaactatt aagatagcaa tcacattcaa tagccaaata
     1741 ctataccacc tgagcgatgc aacgaaatga tcaatttgag caaaaatgct gcatatttag
     1801 gacggcatca ttatagaaat gcttcttgct gtgtactttt ctctcgtctg gcagctgttt
     1861 cgccgttatt gttaaaaccg gcttaagtta ggtgtgtttt ctacgactag tgaatgccct
     1921 actagaagat gtgtgttgca caaaatgtcc ctggaataac caatttgaag tgcagatagc
     1981 agtaaacgta agctaatatg aatattattt aactgtaatg ttttaatatc gctggacatt
     2041 actaataaac ccactataaa cacatgtaca tatgtatgtt ttggcataca atgagtagtt
     2101 ggggaaaaaa tgtgtaaaag caccgtgacc atcacagcat aaagataacc agctgaagta
     2161 tcgaatatga gtaaccccca aattgaatca catgccgcaa ctgataggac ccatggaagt
     2221 acactcttca tggcgatata caagacacac acaagcacga acacccagtt gcggaggaaa
     2281 ttctccgtaa atgaaaaccc aatcggcgaa caattcatac ccatatatgg taaaagtttt
     2341 gaacgcgact tgagagcgga gagcattgcg gctgataagg ttttagcgct aagcgggctt
     2401 tataaaacgg gctgcgggac cagttttcat atcactaccg tttgagttct tgtgctgtgt
     2461 ggatactcct cccgacacaa agccgctcca tcagccagca gtcgtctaat ccagagaccc
     2521 cggatcgggg taccatcatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     2581 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     2641 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     2701 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     2761 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     2821 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     2881 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     2941 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     3001 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     3061 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     3121 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     3181 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     3241 acgagctgta caagccaagc ttgaattctg cagattctga tgtgaagcag accatatacg
     3301 aagttaataa gtacgccaag cgatccaagc taatagaaat cctttccgag caagcagatg
     3361 gcaccattgt gtttgtggag acaaagcgtg gcgccgacga gttagcttcc ttcctgtcag
     3421 aaaaggagtt tccgacgacc tccattcatg gcgatcgtct ccagagtcaa cgcgagcagg
     3481 ccttgcgtga tttcaagaac ggctctatga aggttctcat agccacttca gtggcttctc
     3541 gtggattaga cattaaaaac atcaagcatg tgatcaacta tgacatgccc agcaagatcg
     3601 atgactacgt acatcgcatt ggacgtacag gtcgtgtagg caataatgga cgagccacaa
     3661 gcttctttga ccctgaaaag gatcgagcta ttgctgcgga cttggtaaaa atcttggagg
     3721 gatctggcca gactgttccg gactttctac gcacctgtgg tgccggcggt gatgggggtt
     3781 actccaatca aaatttcggt ggggtggatg tgcgtggcag gggaaattac gtcggcgatg
     3841 ccaccaatgt cgaggaagaa gagcaatggg attgaatcca gcacagtggc ggccgctcga
     3901 gtctagaggg cccgcggttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt
     3961 ctacgcgtac cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga
     4021 ctgtgccttc taaggcctga gctcgctgat cagcctcgat cgaggatcca gacatgataa
     4081 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     4141 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     4201 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     4261 aaagcaagta aaacctctac aaatgtggta tggctgatta tgatcagtcg accgatgccc
     4321 ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc
     4381 gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg
     4441 gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg
     4501 gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt
     4561 ttcggcgaga agcaggccat tatcgccggc atggcggccg acgcgctggg ctacgtcttg
     4621 ctggcgttcg cgacgcgagg ctggatggcc ttccccatta tgattcttct cgcttccggc
     4681 ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga cgaccatcag
     4741 ggacagcttc aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4801 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4861 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4921 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4981 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5041 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5101 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5161 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5221 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5281 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5341 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5401 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5461 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5521 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5581 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5641 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5701 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5761 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5821 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc
     5881 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5941 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6001 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6061 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6121 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg
     6181 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6241 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6301 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6361 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6421 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6481 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6541 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     6601 atcacgaggc cctttcgtct tcaagaatta attctatatt ctaaaaacac aaatgatact
     6661 tctaa
//
