LOCUS       sequence-407103-        6977 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CAf40 expression in Schneider cells for imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6977)
  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 6977)
  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-mEGFP-CAF40 isoform A (HK51)
FEATURES             Location/Qualifiers
     source          1..6977
                     /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 with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     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             4235..4276
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4286..4303
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4326..4343)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4376..4510
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4400..4419)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4454..4473
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(5111..5699)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5191..5210)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5870..6661,6662..6730))
                     /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     6493..6512
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6731..6835)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6903..6921
                     /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 gtccaagctg agcaaagacc ccaacgagaa
     3181 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     3241 cgagctgtac aagccaagct tgaattctgc agatatgagt gctcaaccga gtccgcatat
     3301 gaatcctcag cagcagcagc agcagcaaca gcagcagcag cagaccgagc aggagaaggt
     3361 gtaccagtgg atcaatgagc tggcccatcc ggacacgcgt gaaaccgctc tgctcgagct
     3421 aagcaagaag cgtgagacgg acctggcccc catgctgtgg aacagcttcg ggaccgcctg
     3481 cgccctgttg caggagattg ttaacatcta cccatcgata acgccgccca ctttgacggc
     3541 ccaccagtcg aaccgcgtgt gcaacgccct ggcgctgctg cagtgcgtcg cctcgcaccc
     3601 ggagactcgc acggccttcc tgcaggccca gataccgctg tacttgtacc ctttcttgtc
     3661 gaccacgtcc aagaccaggc ccttcgagta cttgcgcctg accagtttgg gcgtgattgg
     3721 tgctctggtc aagaccgacg aacaggaggt gatcaccttt ctgctgacca ccgagatcgt
     3781 gcccctttgt ctgagcatca tggacagcgg atcggagctg agcaagactg tggccacttt
     3841 catcatccag aagatcctgc tcgatgagtc gggtctgtcg tacatctgcc agacctacga
     3901 acgcttttcg cacgtggcca tcaccctggg caaaatggtc atccagctgg cgaaggatcc
     3961 atgcgcccgg ctgttgaagc atgtggtgcg ctgctatctg cgtctttcgg acaatacacg
     4021 cgctcgcaag gccctgggac agtgtctgcc cgatcagctg cgtgacggca cctttgcgct
     4081 gtgcctgcaa gaggacaagt cgaccaagca gtggctgcag atgctgctca agaacctgga
     4141 actcggcgcc actccgcagc agatcggcat gtcgccactg ggctccatcc agcacagtgg
     4201 cggccgctcg agtctagagg gcccgcggtt cgaaggtaag cctatcccta accctctcct
     4261 cggtctcgat tctacgcgta ccggtcatca tcaccatcac cattgagttt aaacccgctg
     4321 atcagcctcg actgtgcctt ctaaggcctg agctcgctga tcagcctcga tcgaggatcc
     4381 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     4441 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     4501 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     4561 ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatcagtc
     4621 gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga
     4681 ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg
     4741 cagcgctctg ggtcattttc ggcgaggacc gctttcgctg gagcgcgacg atgatcggcc
     4801 tgtcgcttgc ggtattcgga atcttgcacg ccctcgctca agccttcgtc actggtcccg
     4861 ccaccaaacg tttcggcgag aagcaggcca ttatcgccgg catggcggcc gacgcgctgg
     4921 gctacgtctt gctggcgttc gcgacgcgag gctggatggc cttccccatt atgattcttc
     4981 tcgcttccgg cggcatcggg atgcccgcgt tgcaggccat gctgtccagg caggtagatg
     5041 acgaccatca gggacagctt caaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5101 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5161 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5221 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5281 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5341 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5401 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5461 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5521 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     5581 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5641 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5701 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5761 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5821 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5881 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5941 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6001 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6061 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6121 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6181 ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6241 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6301 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6361 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6421 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6481 cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6541 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6601 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6661 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6721 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6781 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6841 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     6901 aaaataggcg tatcacgagg ccctttcgtc ttcaagaatt aattctatat tctaaaaaca
     6961 caaatgatac ttctaaa
//
