LOCUS       sequence-407109-        7721 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CCR4 expression in Schneider cells for imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7721)
  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 7721)
  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-CCR4 isoform A (HK52)
FEATURES             Location/Qualifiers
     source          1..7721
                     /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(2843..2862)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3190..3211
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4979..5020
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5030..5047
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5070..5087)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5120..5254
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5144..5163)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5198..5217
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(5855..6443)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5935..5954)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6614..7405,7406..7474))
                     /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     7237..7256
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7475..7579)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7647..7665
                     /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 agatatgaaa ggcaatcatt ataaaatgtc
     3301 tcgtaacaaa gacaaatacg acagcgccaa tcggcggcag cagatctttc tgtcgcagga
     3361 ggacatagct gcgggcaaga agacgaactg gagtggccta gagattactg gctgcgtccg
     3421 caatatcagc ccgtcgctgt gggaatttga gcatctaacc gccctctacc tgaacgacaa
     3481 ccagctgctg cgattgcccg cagacgttgg catgctgacc agcctccgaa ctttggacct
     3541 gtccagcaat aagctaagaa gtcttcccgc agagctcggc gagctcatcc agttgcggga
     3601 actgttgctg aacaacaact ttctgcgcgt actgccttac gagatcggca agctgttcca
     3661 cctcgtcata ctcggcctca tgggcaatcc gctgcaaaag gagttcatga acatctacaa
     3721 cgaaccgaac ggcacgcaga aactgctcac ctacatgctg gacaacttgt catttaccgt
     3781 gaatccaccg ccccagaggc cctggctgcc actggccaag cccaacaaaa cgcgaccagc
     3841 ctgcattttt acggtcatgt gctataatgt gctctgcgac aagtacgcga cgcgacaaat
     3901 gtacggatac tgtccgtcgt gggcgctatg ctgggagtac cgaaaaaagt cgattatcga
     3961 cgagatacgg cactatgcag cggacattat cagtctgcag gagatcgaaa cggagcaatt
     4021 ctatcacttc ttcctgccgg aactcaagaa cgatggatac gagggaatct tctcaccgaa
     4081 gtcgcgtgct aagactatgt ccgagctgga gaggaagtac gtcgatggct gtgcgatatt
     4141 cttcagggcg tccaagttta cgctgatcaa ggaatcactg atcgagttca atcagctggc
     4201 aatggccaat gccgagggct ccgacaacat gctgaaccgc gtaatgccta aggataacat
     4261 cggtctggcc gcactgctca aggtgaagga gaacgcatgg gagccgatgt ccgaggtgac
     4321 gcagatctcg cagccgctgc tcgtctgcac ggcgcacata cactgggacc ctgagttctg
     4381 cgacgtcaag ctcatccaga cgatgatgct tagcaatgag ttgaagacga tcatcgacga
     4441 ggcgagccac agtttccgac ctggtcacaa gaacgactcc aatgctgtcc agctgctgct
     4501 gtgcggtgac ttcaactcgc tacccgattc aggcgttgtg gagtttctcg gcaagggccg
     4561 cgtttccatg gatcatttgg acttcaagga catgggctac aagtcctgcc tgcagcggct
     4621 gctctcgaac gacaccaacg agtttacgca ctcgttcaag ctagcctccg cctacaacga
     4681 ggacatcatg ccgcacacca actatacgtt cgattttaag ggcatcatcg actacatttt
     4741 ctacacgaag acgggcatgg tgccgctggg cctgctgggt cctgtctcca atgattggct
     4801 gcgcgagaat aaggttgttg gatgcccaca tccgcatata ccctctgatc acttcccact
     4861 gctggtcgag ctagagctga tgcatacggc tagccaacag gcgcctccta acgggctgat
     4921 caatcgccgg atccagcaca gtggcggccg ctcgagtcta gagggcccgc ggttcgaagg
     4981 taagcctatc cctaaccctc tcctcggtct cgattctacg cgtaccggtc atcatcacca
     5041 tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctaagg cctgagctcg
     5101 ctgatcagcc tcgatcgagg atccagacat gataagatac attgatgagt ttggacaaac
     5161 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     5221 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     5281 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     5341 tggtatggct gattatgatc agtcgaccga tgcccttgag agccttcaac ccagtcagct
     5401 ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca
     5461 tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat tttcggcgag gaccgctttc
     5521 gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg
     5581 ctcaagcctt cgtcactggt cccgccacca aacgtttcgg cgagaagcag gccattatcg
     5641 ccggcatggc ggccgacgcg ctgggctacg tcttgctggc gttcgcgacg cgaggctgga
     5701 tggccttccc cattatgatt cttctcgctt ccggcggcat cgggatgccc gcgttgcagg
     5761 ccatgctgtc caggcaggta gatgacgacc atcagggaca gcttcaaggc cagcaaaagg
     5821 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5881 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5941 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6001 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6061 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6121 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6181 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6241 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     6301 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6361 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6421 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6481 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6541 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6601 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6661 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6721 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6781 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6841 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6901 atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg
     6961 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7021 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7081 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7141 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7201 ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa
     7261 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7321 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7381 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7441 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7501 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7561 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     7621 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcaag
     7681 aattaattct atattctaaa aacacaaatg atacttctaa a
//
