LOCUS       sequence-401685-        6689 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 6689)
  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 6689)
  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..6689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..2520
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2540..2542,2543..2545,2546..3256)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2585..2606)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2846..2865)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3193..3214
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3926..3967
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3977..3994
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4017..4034)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4067..4201
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4091..4110)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4145..4164
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4823..5411)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4903..4922)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5582..6373,6374..6442))
                     /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     6205..6224
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6443..6547)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6615..6633
                     /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 atatcaatat
      181 gtacattcga gaaagaagcg atcaaagaag cgtcttcggg cgagtaggag aatgcggagg
      241 agaaggagaa cgagctgatc tagtatctct ccacaatcca atgccaactg accaactggc
      301 catattcgga gcaatttgaa gccaatttcc atcgcctggc gatcgctcca ttcttggcta
      361 tatgtttttc accgttcccg gggccatttt caaagactcg tcggtaagat aagattgtgt
      421 cactcgctgt ctctcttcat ttgtcgaaga atgctgagga atttcgcgat gacgtcggcg
      481 agtattttga agaatgagaa taatttgtat ttatacgaaa atcagttagt ggaattttct
      541 acaaaaacat gttatctata gataattttg ttgcaaaata tgttgactat gacaaagatt
      601 gtatgtatat acctttaatg tattctcatt ttcttatgta tttataatgg caatgatgat
      661 actgatgata ttttaagatg atgccagacc acaggctgat ttctgcgtct tttgccgaac
      721 gcagtgcatg tgcggttgtt gttttttgga atagtttcaa ttttcggact gtccgctttg
      781 atttcagttt cttggcttat tcaaaaagca aagtaaagcc aaaaaagcga gatggcaata
      841 ccaaatgcgg caaaacggta gtggaaggaa aggggtgcgg ggcagcggaa ggaagggtgg
      901 ggcggggcgt ggcggggtct gtggctgggc gcgacgtcac cgacgttgga gccactcctt
      961 tgaccatgtg tgcgtgtgtg tattattcgt gtctcgccac tcgccggttg tttttttctt
     1021 tttatctcgc tctctctagc gccatctcgt acgcatgctc aacgcaccgc atgttgccgt
     1081 gtcctttatg cgtcattttg gctcgaaata ggcaattatt taaacaaaga ttagtcaacg
     1141 aaaacgctaa aataaataag tctacaatat ggttacttat tgccatgtgt gtgcagccaa
     1201 cgatagcaac aaaagcaaca acacagtggc tttccctctt tcactttttg tttgcaagcg
     1261 cgtgcgagca agacggcacg accggcaaac gcaattacgc tgacaaagag cagacgaagt
     1321 tttggccgaa aaacatcaag gcgcctgata cgaatgcatt tgcaataaca attgcgatat
     1381 ttaatattgt ttatgaagct gtttgacttc aaaacacaca aaaaaaaaaa taaaacaaat
     1441 tatttgaaag agaattagga atcggacagc ttatcgttac gggctaacag cacaccgaga
     1501 cgaaatagct tacctgacgt cacagcctct ggaagaactg ccgccaagca gagagagaga
     1561 gaaaaagagg gagagcagct tagaccgcat gtgcttgtgt gtgaggcgtc tctctcttcg
     1621 tctcctgttt gcgcaaacgc atagactgca ctgagaaaat cgattaccta ttttttatga
     1681 atgaatattt gcactattac tattcaaaac tattaagata gcaatcacat tcaatagcca
     1741 aatactatac cacctgagcg atgcaacgaa atgatcaatt tgagcaaaaa tgctgcatat
     1801 ttaggacggc atcattatag aaatgcttct tgctgtgtac ttttctctcg tctggcagct
     1861 gtttcgccgt tattgttaaa accggcttaa gttaggtgtg ttttctacga ctagtgatgc
     1921 ccctactaga agatgtgtgt tgcacaaatg tccctgaata accaatttga agtgcagata
     1981 gcagtaaacg taagctaata tgaatattat ttaactgtaa tgttttaata tcgctggaca
     2041 ttactaataa acccactata aacacatgta catatgtatg ttttggcata caatgagtag
     2101 ttggggaaaa aatgtgtaaa agcaccgtga ccatcacagc ataaagataa ccagctgaag
     2161 tatcgaatat gagtaacccc caaattgaat cacatgccgc aactgatagg acccatggaa
     2221 gtacactctt catggcgata tacaagacac acacaagcac gaacacccag ttgcggagga
     2281 aattctccgt aaatgaaaac ccaatcggcg aacaattcat acccatatat ggtaaaagtt
     2341 ttgaacgcga cttgagagcg gagagcattg cggctgataa ggttttagcg ctaagcgggc
     2401 tttataaaac gggctgcggg accagttttc atatcactac cgtttgagtt cttgtgctgt
     2461 gtggatactc ctcccgacac aaagccgctc catcagccag cagtcgtcta atccagagac
     2521 cccggatcgg ggtaccaaca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     2581 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     2641 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     2701 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     2761 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     2821 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     2881 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     2941 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     3001 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     3061 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     3121 gctgctgccc gacaaccact acctgagcac ccagtccaag ctgagcaaag accccaacga
     3181 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     3241 ggacgagctg tacaagccaa gcttgaattc tgcagattct gatgtgaagc agaccatata
     3301 cgaagttaat aagtacgcca agcgatccaa gctaatagaa atcctttccg agcaagcaga
     3361 tggcaccatt gtgtttgtgg agacaaagcg tggcgccgac gagttagctt ccttcctgtc
     3421 agaaaaggag tttccgacga cctccattca tggcgatcgt ctccagagtc aacgcgagca
     3481 ggccttgcgt gatttcaaga acggctctat gaaggttctc atagccactt cagtggcttc
     3541 tcgtggatta gacattaaaa acatcaagca tgtgatcaac tatgacatgc ccagcaagat
     3601 cgatgactac gtacatcgca ttggacgtac aggtcgtgta ggcaataatg gacgagccac
     3661 aagcttcttt gaccctgaaa aggatcgagc tattgctgcg gacttggtaa aaatcttgga
     3721 gggatctggc cagactgttc cggactttct acgcacctgt ggtgccggcg gtgatggggg
     3781 ttactccaat caaaatttcg gtggggtgga tgtgcgtggc aggggaaatt acgtcggcga
     3841 tgccaccaat gtcgaggaag aagagcaatg ggattgaatc cagcacagtg gcggccgctc
     3901 gagtctagag ggcccgcggt tcgaaggtaa gcctatccct aaccctctcc tcggtctcga
     3961 ttctacgcgt accggtcatc atcaccatca ccattgagtt taaacccgct gatcagcctc
     4021 gactgtgcct tctaaggcct gagctcgctg atcagcctcg atcgaggatc cagacatgat
     4081 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     4141 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     4201 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     4261 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatcagt cgaccgatgc
     4321 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg
     4381 ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct
     4441 gggtcatttt cggcgaggac cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg
     4501 cggtattcgg aatcttgcac gccctcgctc aagccttcgt cactggtccc gccaccaaac
     4561 gtttcggcga gaagcaggcc attatcgccg gcatggcggc cgacgcgctg ggctacgtct
     4621 tgctggcgtt cgcgacgcga ggctggatgg ccttccccat tatgattctt ctcgcttccg
     4681 gcggcatcgg gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat gacgaccatc
     4741 agggacagct tcaaggatcg ctcgcggctc ttaccagcca gcaaaaggcc aggaaccgta
     4801 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4861 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4921 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4981 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca
     5041 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5101 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5161 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5221 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     5281 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5341 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5401 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5461 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5521 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5581 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5641 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5701 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5761 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5821 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5881 acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5941 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6001 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6061 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6121 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6181 gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6241 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6301 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6361 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6421 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6481 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6541 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     6601 cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcaagga attaattcta
     6661 tattctaaaa acacaaatga tacttctaa
//
