LOCUS       sequence-420339-        8527 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of an ActA FKBP payload for MinDE recruitment circuits in
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8527)
  AUTHORS   Rajasekaran R, Chang CC, Weix EWZ, Galateo TM, Coyle SM
  TITLE     A programmable reaction-diffusion system for spatiotemporal cell 
            signaling circuit design.
  JOURNAL   Cell. 2023 Dec 27:S0092-8674(23)01339-9. doi: 
            10.1016/j.cell.2023.12.007.
  PUBMED    38181787
REFERENCE   2  (bases 1 to 8527)
  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: "Cell. 2023 
            Dec 27:S0092-8674(23)01339-9. doi: 10.1016/j.cell.2023.12.007."
COMMENT     Sequence Label: pTwist_Lenti_SFFV-ActA(1-584)-BFP-FKBP
FEATURES             Location/Qualifiers
     source          1..8527
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             249..293
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    560..677
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        773..1174
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      1225..1234
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3013..3711
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             3745..4068
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     LTR             4211..4444
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     rep_origin      4705..4840
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      complement(4958..5546)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5038..5057)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(5908..5976,5977..6768)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6126..6145)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6781..6917
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     enhancer        6994..7297
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7298..7501
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7452..7472
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             7516..7696
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7740..7865
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    join(8358..8527,1..64)
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc
       61 tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg
      121 ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg
      181 acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc
      241 agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt
      301 ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct
      361 tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat
      421 attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag
      481 gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat
      541 ctcgacggta tcggttaact tttaaaagaa aaggggggat tggggggtac agtgcagggg
      601 aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta
      661 caaaaattca aaattttggc tcccgatcgt tgcgttacac acacaattac tgctgatcga
      721 gtgtagcctt cgaatgaaag accccacctg taggtttggc aagatagctg cagtaacgcc
      781 attttgcaag gcatggaaaa ataccaaacc aagaatagag aagttcagat caagggcggg
      841 tacatgaaaa tagctaacgt tgggccaaac aggatatctg cggtgagcag tttcggcccc
      901 ggcccggggc caagaacaga tggtcaccgc agtttcggcc ccggcccgag gccaagaaca
      961 gatggtcccc agatatggcc caaccctcag cagtttctta agacccatca gatgtttcca
     1021 ggctccccca aggacctgaa atgaccctgc gccttatttg aattaaccaa tcagcctgct
     1081 tctcgcttct gttcgcgcgc ttctgcttcc cgagctctat aaaagagctc acaacccctc
     1141 actcggcgcg ccagtcctcc gattgactga gtcgccctga tcattgtcga tcctaccatc
     1201 cactcgacac acccgccagg atccgccacc atgggactga acagatttat gcgggccatg
     1261 atggtggttt tcattactgc caactgcatt acgattaacc ccgacataat attcgcagcg
     1321 actgacagcg aagattctag tttgaacaca gacgaatggg aggaggaaaa gacagaagag
     1381 cagcccagcg aggtgaatac cggaccacgc tacgaaactg cacgtgaggt gagttcacgg
     1441 gatattaaag aactcgagaa aagcaataag gtgagaaaca ctaacaaagc agacctaatc
     1501 gcaatgttga aagaaaaagc cgagaaaggt ccaaacatca ataataacaa cagtgaacag
     1561 actgagaatg ccgctataaa tgaagaggct tcaggagccg accgacccgc tatccaggtg
     1621 gagcgtcggc accctggcct gccctccgac agcgccgccg aaattaaaaa gaggaggaaa
     1681 gccatcgcat cctcggatag tgagcttgaa agcctgactt atccggataa accaacaaaa
     1741 gtaaataaga agaaggtggc gaaggagtca gttgccgatg cttctgaaag tgacttagac
     1801 tctagcatgc agtcagcaga cgagtcttcc ccacaacctc tgaaagccaa ccaacagccc
     1861 ttcttcccta aggtctttaa aaaaataaag gatgccggga aatgggtccg cgacaagatc
     1921 gacgaaaatc ctgaggtcaa gaaggcgatt gttgataaat ccgccgggct catcgaccag
     1981 ctgctcacca agaagaagag tgaggaggtg aatgcttctg acttcccgcc cccacctacg
     2041 gatgaagagc tgcgactggc tttgccagag acccctatgc ttctggggtt taatgctcct
     2101 gccacatcag aaccgagctc attcgagttt ccaccacccc ctaccgacga agagctgagg
     2161 cttgctttgc ccgagacgcc aatgctgctt ggttttaatg cccctgccac ctccgagccc
     2221 agctcgttcg aattcccacc tcctcccact gaggatgaac tcgagatcat ccgggagaca
     2281 gcatcctctc tcgattcttc ctttacccgc ggggacttag cttccttgcg aaatgctatt
     2341 aaccgccatt cccaaaattt ctctgacttc ccacctatcc ccacagaaga ggagttgaac
     2401 gggaggggcg gcagaccaac atctgaggag ttctcctcgc tgaatagtgg tgattttacc
     2461 gatgacgaga acagcgagac aacagaagaa gaaattgatc gcctagccga tctgagagat
     2521 agaggaacag gaaaacactc aaggaatgcg ggatttctgc ccctgaatcc gttcgccagc
     2581 agcccggttc cttctttaag tcccaaggtc tccaaaatca gcgcaccggc tctgatcagt
     2641 gacataacta aaaagacgcc atttaagaat ccttcccagc ccctgaatgt gtttaataag
     2701 aagactacca ctaagaccgt gactaagaaa cctacccctg tgaagaccgc cccaaagcta
     2761 gcagaacttc ctgccacaaa gccccaggag accgtactta gggaaaacaa gacccccttt
     2821 atagagaagc aggcagagac aaacaagcag tcaatcaaca tgcccagcct ccccgtcatc
     2881 cagaaagagg ctaccgagag cgataaggag gagatgaaac cacaaaccga ggaaaagatg
     2941 gtagaggaaa gcgaatcagc caacaacgca aacggcaaga acggtagcgg cggaagcggt
     3001 ggcggtggca gcatgagcga gctgattaag gagaacatgc acatgaagct gtacatggag
     3061 ggcaccgtgg acaaccatca cttcaagtgc acatccgagg gcgaaggcaa gccctacgag
     3121 ggcacccaga ccatgagaat caaggtggtc gagggcggcc ctctcccctt cgccttcgac
     3181 atcctggcta ctagcttcct ctacggcagc aagaccttca tcaaccacac ccagggcatc
     3241 cccgacttct tcaagcagtc cttccctgag ggcttcacat gggagagagt caccacatac
     3301 gaagacgggg gcgtgctgac cgctacccag gacaccagcc tccaggacgg ctgcctcatc
     3361 tacaacgtca agatcagagg ggtgaacttc acatccaacg gccctgtgat gcagaagaaa
     3421 acactcggct gggaggcctt caccgagacg ctgtaccccg ctgacggcgg cctggaaggc
     3481 agaaacgaca tggccctgaa gctcgtgggc gggagccatc tgatcgcaaa catcaagacc
     3541 acatatagat ccaagaaacc cgctaagaac ctcaagatgc ctggcgtcta ctatgtggac
     3601 tacagactgg aaagaatcaa ggaggccaac aacgagacct acgtcgagca gcacgaggtg
     3661 gcagtggcca gatactgcga cctccctagc aaactggggc acaagcttaa tgacagcgcc
     3721 ggaagtgctg gctctgcggg taccggagtg caggtggaaa ccatctcccc aggagacggg
     3781 cgcaccttcc ccaagcgcgg ccagacctgc gtggtgcact acaccgggat gcttgaagat
     3841 ggaaagaaat ttgattcctc ccgggacaga aacaagccct ttaagtttat gctaggcaag
     3901 caggaggtga tccgaggctg ggaagaaggg gttgcccaga tgagtgtggg tcagagagcc
     3961 aaactgacta tatctccaga ttatgcctat ggtgccactg ggcacccagg catcatccca
     4021 ccacatgcca ctctcgtctt cgatgtggag cttctaaaac tggaatgact cgagggcccg
     4081 catccaccat cgcagactta tcatcttatt ccagatgcgt gcggatggaa ttcgagctcg
     4141 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     4201 aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt
     4261 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     4321 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     4381 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     4441 agcagtcctg gccaacgtga gcaccgtgct gacctccaaa tatcgttaag ctggagcctg
     4501 ggagccggcc tggccctccg ccccccccac ccccgcagcc cacccctggt ctttgaataa
     4561 agtctgagtg agtggccgac agtgcccgtg gagttctcgt gacctgaggt gcagggccgg
     4621 cgctagggac acgtccgtgc acgtgccgag gccccctgtg cagctgcaag ggacaggcct
     4681 agccctgcag gcctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4741 ccgcctcatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     4801 gagctattcc agaagtagtg aggacgcttt tttggaggcc gaggcttttg caaagatcga
     4861 acaagagaca ggacctgcag gttaattaaa tttaaatcat gtgagcaaaa ggccagcaaa
     4921 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4981 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5041 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5101 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5161 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5221 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5281 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5341 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5401 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5461 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5521 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5581 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5641 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5701 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5761 tatttcgttc atccatagtt gcatttaaat ggccggcctg gcgcgccgtt taaacctaga
     5821 tattgatagt ctgatcggtc aacgtataat cgagtcctag cttttgcaaa catctatcaa
     5881 gagacaggat cagcaggagg ctttcgcatg agtattcaac atttccgtgt cgcccttatt
     5941 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     6001 aaagatgctg aagatcagtt gggtgcgcga gtgggttaca tcgaactgga tctcaacagc
     6061 ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc caatgatgag cacttttaaa
     6121 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     6181 cgcatacact attctcagaa tgacttggtt gagtattcac cagtcacaga aaagcatctt
     6241 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6301 gcggccaact tacttctgac aacgattgga ggaccgaagg agctaaccgc ttttttgcac
     6361 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6421 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacctt gcgtaaacta
     6481 ttaactggcg aactacttac tctagcttcc cggcaacagt tgatagactg gatggaggcg
     6541 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6601 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6661 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6721 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaacc gattctaggt
     6781 gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc
     6841 cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac
     6901 cagaaattta tccttaagat cccgaatcgt ttaaacgcga tcgcagtaat caattacggg
     6961 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     7021 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     7081 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     7141 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     7201 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     7261 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
     7321 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     7381 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     7441 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     7501 tcgtttagtg aaccggggtc tctctggtta gaccagatct gagcctggga gctctctggc
     7561 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     7621 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     7681 tggaaaatct ctagcagtgg cgcccgaaca gggacctgaa agcgaaaggg aaaccagagc
     7741 tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac
     7801 tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga
     7861 gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag
     7921 ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat
     7981 tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc
     8041 tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa
     8101 ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga
     8161 tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga
     8221 cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta
     8281 aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa
     8341 aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact
     8401 atgggcgcag cctcaatgac gctgacggta caggccagac aattattgtc tggtatagtg
     8461 cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca
     8521 gtctggg
//
