LOCUS       sequence-467429-        6490 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AC5 promoter driven FRB-MCP for drosophila cell expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6490)
  AUTHORS   Zhang X, Mille-Fragoso LS, Kaseniit KE, Lee AP, Zhang M, Call CC, Hu
            Y, Xie Y, Gao XJ
  TITLE     Post-transcriptional modular synthetic receptors.
  JOURNAL   Nat Chem Biol. 2025 Mar 28. doi: 10.1038/s41589-025-01872-w.
  PUBMED    40155716
REFERENCE   2  (bases 1 to 6490)
  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 Chem 
            Biol. 2025 Mar 28. doi: 10.1038/s41589-025-01872-w."
COMMENT     Sequence Label: APL0041 Ac5-FRB-MCP
FEATURES             Location/Qualifiers
     source          1..6490
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     4..22
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        86..2595
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     regulatory      2648..2657
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3826..3867
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3877..3894
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3917..3934)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3967..4101
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3991..4010)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4045..4064
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4702..5290)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4782..4801)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5461..6252,6253..6321))
                     /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     6084..6103
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6322..6426)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaaaataggc gtatcacgag gccctttcgt cttcaagaat taattctata ttctaaaaac
       61 acaaatgata cttctaaaaa aaaatcatga atggcatcaa ctctgaatca aatctttgca
      121 gatgcaccta cttctcattt ccactgtcac atcatttttc cagatctcgc tgcctgttat
      181 gtggcccaca aaccaagaca cgttttatgg ccattaaagc tggctgatcg tcgccaaaca
      241 ccaaatacat aatgaatatg tacacattcg agaaagaagc gatcaaagaa gcgtcttcgg
      301 gcggagtagg agaatgcgga ggagaaggag aacgagctga tctagtatct ctccacaatc
      361 caatgccaac tgaccaactg gccatattcg gagcaatttg aagccaattt ccatcgcctg
      421 gcgatcgctc cattcttggc tatatgtttt tcaccgttac ccggggccat tttcaaagac
      481 tcgtcggcaa gataagattg tgtcactcgc tgtctctctt catttgtcga agaatgctga
      541 ggaatttcgc gatgacgtcg gcgagtattt tgaagaatga gaataatttg tatttatacg
      601 aaaatcagtt agtggaattt tctacaaaaa catgttatct atagataatt ttgttgcaaa
      661 atatgttgac tatgacaaag attgtatgta tataccttta atgtattctc attttcttat
      721 gtatttataa tggcaatgat gatactgatg atattttaag atgatgccag accaaaaggc
      781 ttgaatttct gcgtcttttg ccgaacgcag tgcatgtgca attgttgttt tttggaatat
      841 tcaattttcg gactgtccgc tttgatttca gtttcttggc ttattcaaaa agcaaagtaa
      901 agccaaaaaa gcgagatggc aataccaaat gcggcaaaac ggtagtggaa ggaaaggggt
      961 gcggggcagc ggaaggaagg gtggggcggg gcgtggcggg gtctgtggct gggcgcgacg
     1021 tcaccgacgt tggagccact cctttgacca tgtgtgcgtg tgtgtattat tcgtgtctcg
     1081 ccactcgccg gttgtttttt tctttttatg ctgcgctctc tctagcgcca tctcgcttac
     1141 gcatgctcaa cgcaccgcat gttgccgttt ccttttatgc gtcattttgg ctcgaaatag
     1201 gcaattattt aaacaaagat tagtcaacga aaacgctaaa ataaataagt ctacaatatg
     1261 gttacttatt gccatgtgtg tgcagccaac gatagcaaca aaagcaacaa cacaggtggc
     1321 tttccctctt tcactttttg tttgcaagcc gcgtgcgagc aagacggcac gaccggcaaa
     1381 cgcaattacg ctgacaaaga gcagacgaag ttttggcgaa aaacatcaag gcgcctgata
     1441 cgaatgcatt tgcaataaca attgcgatat ttaatattgt ttatgaagct gtttgacttc
     1501 aaaacacaca aaaaaaaaaa taaaacaaat tatttgaaag agaattagga atcggacgct
     1561 tatcgttagg gtaacaacaa gaaatgctta ctgagtcaca gcctctggaa aactgccgca
     1621 agccagagag agagagaaaa agagggagag cagcttagac cgcatgtgct tgtgtgtgag
     1681 gcgtctctct cttcgtctct gttgcgcaaa cgcatagact gcactgaaaa aatcgattac
     1741 ctatttttta tgaatgaata tttgcactat tactattcaa aactattaag atagcaatca
     1801 cattcaatag ccaaatacta taccacctga gcgatgcaac gaaatgatca atttgagcaa
     1861 aaatgctgca tatttaggac ggcatcatta tagaaatgct tcttgctgtg tacttttctc
     1921 tcgtctggca gctgtttcgc cgttattgtt aaaaccggct taagttaggt gtgttttcta
     1981 cgactagtga atgccctact agaagatgtg tgttgcacaa aatgtccctg gaataaccaa
     2041 tttgaagtgc agatagcagt aaacgtaagc taatatgaat attatttaac tgtaatgttt
     2101 taatatcgct ggacattact aataaaccca ctataaacac atgtacatat gtatgttttg
     2161 gcatacaatg agtagttggg gaaaaaatgt gtaaaagcac cgtgaccatc acagcataaa
     2221 gataaccagc tgaagtatcg aatatgagta acccccaaat tgaatcacat gccgcaactg
     2281 ataggaccca tggaagtaca ctcttcatgg cgatatacaa gacacacaca agcacgaaca
     2341 cccagttgcg gaggaaattc tccgtaaatg aaaacccaat cggcgaacaa ttcataccca
     2401 tatatggtaa aagttttgaa cgcgacttga gagcggagag cattgcggct gataaggttt
     2461 tagcgctaag cgggctttat aaaacgggct gcgggaccag ttttcatatc actaccgttt
     2521 gagttcttgt gctgtgtgga tactcctccc gacacaaagc cgctccatca gccagcagtc
     2581 gtctaatcca gagaccccgg atcggggtac ctactagtcc agtgtggtgg aattccgaaa
     2641 tgggatcgcc accatgggat ccctggagat gtggcatgaa ggcctggaag aggcatctcg
     2701 tttgtacttt ggggaaagga acgtgaaagg catgtttgag gtgctggagc ccttgcatgc
     2761 tatgatggaa cggggacccc agactctgaa ggaaacatcc tttaatcagg cctatggtcg
     2821 agatttaatg gaggcccaag agtggtgcag gaagtacatg aaatcaggga atgtcaagga
     2881 cctcctccaa gcctgggacc tctattatca tgtgttccga cgaatctcaa agactagagg
     2941 cgccgggaca ggagggtctg ggggtggcac tggcggatct ggaggagact acaaggacga
     3001 tgacgacaac ggcggatcgg gggggtctgg tggtagtggc ggttcggggg gaggtaccgg
     3061 tatggcgagc aactttacgc aatttgtgtt ggttgacaat ggtggcaccg gagacgtaac
     3121 tgttgcgccg agcaattttg cgaacgggat tgcagaatgg ataagtagta acagccggag
     3181 tcaggcatat aaagtcacat gctcagttcg ccaaagttcc gcacagaacc gaaaatatac
     3241 cataaaggta gaagtgccaa aaggggcgtg gcgatcttat cttaatatgg agcttacgat
     3301 cccaatcttt gcgactaatt cagactgcga gctgattgtc aaggccatgc aaggtctttt
     3361 gaaagacggt aacccgattc cctctgcgat cgccgccaac tcaggaatat atggtggcgg
     3421 aaccggtggc gggtccatgg cctccaattt tacacagttt gtgctcgtgg acaacggcgg
     3481 cactggagac gtcacagtag ctccaagtaa ctttgcgaat gggatagccg agtggatctc
     3541 ttccaattca agatcacaag cctataaggt gacttgtagt gtacgccaat ctagtgccca
     3601 gaaccggaag tataccatta aagtggaggt ccccaaaggt gcatggcgct catacttgaa
     3661 tatggaattg acgattccga tatttgcgac caatagcgac tgcgagctta ttgtaaaagc
     3721 catgcaagga cttctgaaag atggcaaccc gataccttcc gcaatcgctg cgaatagcgg
     3781 aatttactga gcggccgctc gagtctagag ggcccgcggt tcgaaggtaa gcctatccct
     3841 aaccctctcc tcggtctcga ttctacgcgt accggtcatc atcaccatca ccattgagtt
     3901 taaacccgct gatcagcctc gactgtgcct tctaaggcct gagctcgctg atcagcctcg
     3961 atcgaggatc cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     4021 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     4081 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     4141 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tatggctgat
     4201 tatgatcagt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc
     4261 gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg
     4321 acaggtgccg gcagcgctct gggtcatttt cggcgaggac cgctttcgct ggagcgcgac
     4381 gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac gccctcgctc aagccttcgt
     4441 cactggtccc gccaccaaac gtttcggcga gaagcaggcc attatcgccg gcatggcggc
     4501 cgacgcgctg ggctacgtct tgctggcgtt cgcgacgcga ggctggatgg ccttccccat
     4561 tatgattctt ctcgcttccg gcggcatcgg gatgcccgcg ttgcaggcca tgctgtccag
     4621 gcaggtagat gacgaccatc agggacagct tcaaggccag caaaaggcca ggaaccgtaa
     4681 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4741 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4801 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4861 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4921 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4981 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5041 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5101 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     5161 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5221 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5281 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5341 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5401 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5461 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5521 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5581 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5641 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5701 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5761 cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5821 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5881 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5941 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6001 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6061 ctcttgcccg gcgtcaacac gggataatac cgcgccacat agcagaactt taaaagtgct
     6121 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6181 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6241 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6301 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6361 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6421 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     6481 attaacctat
//
