LOCUS       sequence-439133-        6065 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Schneider cell imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6065)
  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 6065)
  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: T5-pAc5.1 EGFP-EcoRV
FEATURES             Location/Qualifiers
     source          1..6065
                     /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             3324..3365
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3375..3392
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3415..3432)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3465..3599
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3489..3508)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3543..3562
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4200..4788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4280..4299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4959..5750,5751..5819))
                     /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     5582..5601
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5820..5924)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5992..6010
                     /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 cagatatcca gcacagtggc ggccgctcga
     3301 gtctagaggg cccgcggttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt
     3361 ctacgcgtac cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga
     3421 ctgtgccttc taaggcctga gctcgctgat cagcctcgat cgaggatcca gacatgataa
     3481 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     3541 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     3601 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     3661 aaagcaagta aaacctctac aaatgtggta tggctgatta tgatcagtcg accgatgccc
     3721 ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc
     3781 gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg
     3841 gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg
     3901 gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt
     3961 ttcggcgaga agcaggccat tatcgccggc atggcggccg acgcgctggg ctacgtcttg
     4021 ctggcgttcg cgacgcgagg ctggatggcc ttccccatta tgattcttct cgcttccggc
     4081 ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga cgaccatcag
     4141 ggacagcttc aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4201 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4261 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4321 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4381 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4441 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4501 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4561 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4621 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4681 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4741 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4801 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4861 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4921 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4981 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5041 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5101 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5161 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5221 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc
     5281 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5341 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5401 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5461 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5521 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg
     5581 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     5641 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     5701 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     5761 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     5821 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5881 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     5941 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     6001 atcacgaggc cctttcgtct tcaagaatta attctatatt ctaaaaacac aaatgatact
     6061 tctaa
//
