LOCUS       sequence-407836-        8130 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 8130)
  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 8130)
  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-CCR4 isoform A-mCherry-PH (EB7)
FEATURES             Location/Qualifiers
     source          1..8130
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..2565
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             4257..4964
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4402..4420)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4670..4689
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             5436..5477
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5487..5504
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5527..5544)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5577..5711
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5601..5620)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5655..5674
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6312..6900)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6392..6411)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7071..7862,7863..7931))
                     /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     7694..7713
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7932..8036)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8104..8122
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cttcaagaat taattctata ttctaaaaac acaaatgata cttctaaaaa aaaatcatga
       61 atggcatcaa ctctgaatca aatctttgca gatgcaccta cttctcattt ccactgtcac
      121 atcatttttc cagatctcgc tgcctgttat gtggcccaca aaccaagaca cgttttatgg
      181 ccattaaagc tggctgatcg tcgccaaaca ccaaatacat aatgaatatg tacacattcg
      241 agaaagaagc gatcaaagaa gcgtcttcgg gcggagtagg agaatgcgga ggagaaggag
      301 aacgagctga tctagtatct ctccacaatc caatgccaac tgaccaactg gccatattcg
      361 gagcaatttg aagccaattt ccatcgcctg gcgatcgctc cattcttggc tatatgtttt
      421 tcaccgttac ccggggccat tttcaaagac tcgtcggcaa gataagattg tgtcactcgc
      481 tgtctctctt catttgtcga agaatgctga ggaatttcgc gatgacgtcg gcgagtattt
      541 tgaagaatga gaataatttg tatttatacg aaaatcagtt agtggaattt tctacaaaaa
      601 catgttatct atagataatt ttgttgcaaa atatgttgac tatgacaaag attgtatgta
      661 tataccttta atgtattctc attttcttat gtatttataa tggcaatgat gatactgatg
      721 atattttaag atgatgccag accaaaaggc ttgaatttct gcgtcttttg ccgaacgcag
      781 tgcatgtgca attgttgttt tttggaatat tcaattttcg gactgtccgc tttgatttca
      841 gtttcttggc ttattcaaaa agcaaagtaa agccaaaaaa gcgagatggc aataccaaat
      901 gcggcaaaac ggtagtggaa ggaaaggggt gcggggcagc ggaaggaagg gtggggcggg
      961 gcgtggcggg gtctgtggct gggcgcgacg tcaccgacgt tggagccact cctttgacca
     1021 tgtgtgcgtg tgtgtattat tcgtgtctcg ccactcgccg gttgtttttt tctttttatg
     1081 ctgcgctctc tctagcgcca tctcgcttac gcatgctcaa cgcaccgcat gttgccgttt
     1141 ccttttatgc gtcattttgg ctcgaaatag gcaattattt aaacaaagat tagtcaacga
     1201 aaacgctaaa ataaataagt ctacaatatg gttacttatt gccatgtgtg tgcagccaac
     1261 gatagcaaca aaagcaacaa cacaggtggc tttccctctt tcactttttg tttgcaagcc
     1321 gcgtgcgagc aagacggcac gaccggcaaa cgcaattacg ctgacaaaga gcagacgaag
     1381 ttttggcgaa aaacatcaag gcgcctgata cgaatgcatt tgcaataaca attgcgatat
     1441 ttaatattgt ttatgaagct gtttgacttc aaaacacaca aaaaaaaaaa taaaacaaat
     1501 tatttgaaag agaattagga atcggacgct tatcgttagg gtaacaacaa gaaatgctta
     1561 ctgagtcaca gcctctggaa aactgccgca agccagagag agagagaaaa agagggagag
     1621 cagcttagac cgcatgtgct tgtgtgtgag gcgtctctct cttcgtctct gttgcgcaaa
     1681 cgcatagact gcactgaaaa aatcgattac ctatttttta tgaatgaata tttgcactat
     1741 tactattcaa aactattaag atagcaatca cattcaatag ccaaatacta taccacctga
     1801 gcgatgcaac gaaatgatca atttgagcaa aaatgctgca tatttaggac ggcatcatta
     1861 tagaaatgct tcttgctgtg tacttttctc tcgtctggca gctgtttcgc cgttattgtt
     1921 aaaaccggct taagttaggt gtgttttcta cgactagtga atgccctact agaagatgtg
     1981 tgttgcacaa aatgtccctg gaataaccaa tttgaagtgc agatagcagt aaacgtaagc
     2041 taatatgaat attatttaac tgtaatgttt taatatcgct ggacattact aataaaccca
     2101 ctataaacac atgtacatat gtatgttttg gcatacaatg agtagttggg gaaaaaatgt
     2161 gtaaaagcac cgtgaccatc acagcataaa gataaccagc tgaagtatcg aatatgagta
     2221 acccccaaat tgaatcacat gccgcaactg ataggaccca tggaagtaca ctcttcatgg
     2281 cgatatacaa gacacacaca agcacgaaca cccagttgcg gaggaaattc tccgtaaatg
     2341 aaaacccaat cggcgaacaa ttcataccca tatatggtaa aagttttgaa cgcgacttga
     2401 gagcggagag cattgcggct gataaggttt tagcgctaag cgggctttat aaaacgggct
     2461 gcgggaccag ttttcatatc actaccgttt gagttcttgt gctgtgtgga tactcctccc
     2521 gacacaaagc cgctccatca gccagcagtc gtctaatcca gagaccccgg atcgggtgca
     2581 tgaaaggcaa tcattataaa atgtctcgta acaaagacaa atacgacagc gccaatcggc
     2641 ggcagcagat ctttctgtcg caggaggaca tagctgcggg caagaagacg aactggagtg
     2701 gcctagagat tactggctgc gtccgcaata tcagcccgtc gctgtgggaa tttgagcatc
     2761 taaccgccct ctacctgaac gacaaccagc tgctgcgatt gcccgcagac gttggcatgc
     2821 tgaccagcct ccgaactttg gacctgtcca gcaataagct aagaagtctt cccgcagagc
     2881 tcggcgagct catccagttg cgggaactgt tgctgaacaa caactttctg cgcgtactgc
     2941 cttacgagat cggcaagctg ttccacctcg tcatactcgg cctcatgggc aatccgctgc
     3001 aaaaggagtt catgaacatc tacaacgaac cgaacggcac gcagaaactg ctcacctaca
     3061 tgctggacaa cttgtcattt accgtgaatc caccgcccca gaggccctgg ctgccactgg
     3121 ccaagcccaa caaaacgcga ccagcctgca tttttacggt catgtgctat aatgtgctct
     3181 gcgacaagta cgcgacgcga caaatgtacg gatactgtcc gtcgtgggcg ctatgctggg
     3241 agtaccgaaa aaagtcgatt atcgacgaga tacggcacta tgcagcggac attatcagtc
     3301 tgcaggagat cgaaacggag caattctatc acttcttcct gccggaactc aagaacgatg
     3361 gatacgaggg aatcttctca ccgaagtcgc gtgctaagac tatgtccgag ctggagagga
     3421 agtacgtcga tggctgtgcg atattcttca gggcgtccaa gtttacgctg atcaaggaat
     3481 cactgatcga gttcaatcag ctggcaatgg ccaatgccga gggctccgac aacatgctga
     3541 accgcgtaat gcctaaggat aacatcggtc tggccgcact gctcaaggtg aaggagaacg
     3601 catgggagcc gatgtccgag gtgacgcaga tctcgcagcc gctgctcgtc tgcacggcgc
     3661 acatacactg ggaccctgag ttctgcgacg tcaagctcat ccagacgatg atgcttagca
     3721 atgagttgaa gacgatcatc gacgaggcga gccacagttt ccgacctggt cacaagaacg
     3781 actccaatgc tgtccagctg ctgctgtgcg gtgacttcaa ctcgctaccc gattcaggcg
     3841 ttgtggagtt tctcggcaag ggccgcgttt ccatggatca tttggacttc aaggacatgg
     3901 gctacaagtc ctgcctgcag cggctgctct cgaacgacac caacgagttt acgcactcgt
     3961 tcaagctagc ctccgcctac aacgaggaca tcatgccgca caccaactat acgttcgatt
     4021 ttaagggcat catcgactac attttctaca cgaagacggg catggtgccg ctgggcctgc
     4081 tgggtcctgt ctccaatgat tggctgcgcg agaataaggt tgttggatgc ccacatccgc
     4141 atataccctc tgatcacttc ccactgctgg tcgagctaga gctgatgcat acggctagcc
     4201 aacaggcgcc tcctaacggg ctgatcaatc gccgggcatc cagcggcggt accaacatgg
     4261 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
     4321 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     4381 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
     4441 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
     4501 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     4561 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg
     4621 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
     4681 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
     4741 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     4801 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     4861 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     4921 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagccaagc ttgaattctg
     4981 cagatggctc catgcacggt ctccaggatg atccagatct gcaggctctg ctgaaaggtt
     5041 ctcagctcct caaagtcaaa agcagctcct ggcgtcgtga acgcttctac aaactgcagg
     5101 aggactgcaa aaccatctgg caggaatccc gtaaagtaat gcgctcccca gagtctcagc
     5161 tcttcagcat tgaggacatt caggaggtac gtatgggtca ccgtaccgaa ggtctggaga
     5221 aattcgctcg cgatatccca gaagatcgct gcttctctat cgtcttcaaa gaccagcgca
     5281 acaccctgga tctgattgct ccatctccag cagatgctca gcactgggtt cagggtctcc
     5341 gcaaaatcat tcaccactcc ggttctatgg accagcgtca gaaataaatc cagcacagtg
     5401 gcggccgctc gagtctagag ggcccgcggt tcgaaggtaa gcctatccct aaccctctcc
     5461 tcggtctcga ttctacgcgt accggtcatc atcaccatca ccattgagtt taaacccgct
     5521 gatcagcctc gactgtgcct tctaaggcct gagctcgctg atcagcctcg atcgaggatc
     5581 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     5641 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     5701 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     5761 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatcagt
     5821 cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg
     5881 actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg
     5941 gcagcgctct gggtcatttt cggcgaggac cgctttcgct ggagcgcgac gatgatcggc
     6001 ctgtcgcttg cggtattcgg aatcttgcac gccctcgctc aagccttcgt cactggtccc
     6061 gccaccaaac gtttcggcga gaagcaggcc attatcgccg gcatggcggc cgacgcgctg
     6121 ggctacgtct tgctggcgtt cgcgacgcga ggctggatgg ccttccccat tatgattctt
     6181 ctcgcttccg gcggcatcgg gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat
     6241 gacgaccatc agggacagct tcaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6301 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6361 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6421 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6481 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6541 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6601 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6661 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6721 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     6781 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6841 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6901 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6961 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7021 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     7081 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     7141 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7201 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7261 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7321 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7381 attgctgcag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7441 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     7501 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7561 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7621 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7681 gcgtcaacac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7741 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7801 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7861 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7921 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7981 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     8041 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     8101 aaaaataggc gtatcacgag gccctttcgt
//
