LOCUS       sequence-466383-        6313 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRB fused with MCP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6313)
  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 6313)
  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: EK0329 CMV-TO-FRB-MCP (FLP-IN)
FEATURES             Location/Qualifiers
     source          1..6313
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(17..220)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(46..66)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        221..600
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     767..786
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        861..965
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(966..1034,1035..1826)
                     /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(1184..1203)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1997..2585
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2486..2505
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2739..2756
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2873..2894
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2909..2914,2915..2932,2933..2939)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2947..2963
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2952..2974
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2971..2987
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2971..2987
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3036..3157
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3047..3066)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3101..3120
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3287..4309)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             complement(3287..4309)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     protein_bind    complement(4317..4364)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     4496..4515
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    4687..4911
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4900..4917
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(4973..5002)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      6154..6163
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6255..6279)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    6289..6307
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    join(6310..6313,1..15)
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 tatcactgat agggagagct ctgcttatat agacctccca ccgtacacgc ctaccgccca
       61 tttgcgtcaa tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc
      121 aaaacaaact cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc
      181 gctatccacg cccattgatg tactgccaaa accgcatcac catggtaata gcgatgacta
      241 atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc
      301 aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt
      361 gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag
      421 tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt
      481 tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg
      541 ggctatgaac taatgacccc gtaattgatt actattaata actagtcaat aatcaatgtc
      601 aacgcgtata tctggcccgt acgaagcagc gcaaaacgcc taaccctaag cagattcttc
      661 atgcaattgt cggtcaagcc ttgccttgtt gtagcttaaa ttttgctcgc gcactactca
      721 gcgacctcca acacacaagc agggagcaga tactggctta actatgcggc atcagagcag
      781 attgtactga gagtgcacca taggggatcg ggagatctcc cgatccgtcg acgtcaggtg
      841 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
      901 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      961 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     1021 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     1081 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     1141 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     1201 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     1261 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     1321 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     1381 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     1441 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     1501 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     1561 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     1621 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     1681 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     1741 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     1801 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1861 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1921 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1981 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     2041 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     2101 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     2161 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     2221 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     2281 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     2341 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     2401 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     2461 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     2521 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     2581 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     2641 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     2701 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     2761 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     2821 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     2881 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     2941 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     3001 agctctagct agaggtcgac ggtatacaga catgataaga tacattgatg agtttggaca
     3061 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     3121 tttatttgta accattataa gctgcaataa acaagttggg gtgggcgaag aactccagca
     3181 tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca
     3241 acctttcata gaaggcggcg gtggaatcga aatctcgtag tacgtgctat tcctttgccc
     3301 tcggacgagt gctggggcgt cggtttccac tatcggcgag tacttctaca cagccatcgg
     3361 tccagacggc cgcgcttctg cgggcgattt gtgtacgccc gacagtcccg gctccggatc
     3421 ggacgattgc gtcgcatcga ccctgcgccc aagctgcatc atcgaaattg ccgtcaacca
     3481 agctctgata gagttggtca agaccaatgc ggagcatata cgcccggagc cgcggcgatc
     3541 ctgcaagctc cggatgcctc cgctcgaagt agcgcgtctg ctgctccata caagccaacc
     3601 acggcctcca gaagaagatg ttggcgacct cgtattggga atccccgaac atcgcctcgc
     3661 tccagtcaat gaccgctgtt atgcggccat tgtccgtcag gacattgttg gagccgaaat
     3721 ccgcgtgcac gaggtgccgg acttcggggc agtcctcggc ccaaagcatc agctcatcga
     3781 gagcctgcgc gacggacgca ctgacggtgt cgtccatcac agtttgccag tgatacacat
     3841 ggggatcagc aatcgcgcat atgaaatcac gccatgtagt gtattgaccg attccttgcg
     3901 gtccgaatgg gccgaacccg ctcgtctggc taagatcggc cgcagcgatc gcatccatgg
     3961 cctccgcgac cggctgcaga acagcgggca gttcggtttc aggcaggtct tgcaacgtga
     4021 caccctgtgc acggcgggag atgcaatagg tcaggctctc gctgaattcc ccaatgtcaa
     4081 gcacttccgg aatcgggagc gcggccgatg caaagtgccg ataaacataa cgatctttgt
     4141 agaaaccatc ggcgcagcta tttacccgca ggacatatcc acgccctcct acatcgaagc
     4201 tgaaagcacg agattcttcg ccctccgaga gctgcatcag gtcggagacg ctgtcgaact
     4261 tttcgatcag aaacttctcg acagacgtcg cggtgagttc aggctttttg gccaaggaag
     4321 ttcctatact ttctagagaa taggaacttc ggaataggaa cttctaggta cgtgaaccat
     4381 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     4441 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     4501 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     4561 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtggggatac cccctagagc
     4621 cccagctggt tctttccgcc tcagaagcca tagagcccac cgcatcccca gaacatcgat
     4681 agatctccat agagcccacc gcatccccag catgcctgct attgtcttcc caatcctccc
     4741 ccttgctgtc ctgccccacc ccacccccca gaatagaatg acacctactc agacaatgcg
     4801 atgcaatttc ctcattttat taggaaagga cagtgggagt ggcaccttcc agggtcaagg
     4861 aaggcacggg ggaggggcaa acaacagatg gctggcaact agaaggcaca gtcgaggctg
     4921 atcagcgggt ttaaactcaa tgatgattat gatgatggtc gacggcgcta ttcagatcct
     4981 cttctgagat gagtttttgt tcgggccctc ctcgagcggc tcagtaaatt ccgctattcg
     5041 cagcgattgc ggaaggtatc gggttgccat ctttcagaag tccttgcatg gcttttacaa
     5101 taagctcgca gtcgctattg gtcgcaaata tcggaatcgt caattccata ttcaagtatg
     5161 agcgccatgc acctttgggg acctccactt taatggtata cttccggttc tgggcactag
     5221 attggcgtac actacaagtc accttatagg cttgtgatct tgaattggaa gagatccact
     5281 cggctatccc attcgcaaag ttacttggag ctactgtgac gtctccagtg ccgccgttgt
     5341 ccacgagcac aaactgtgta aaattggagg ccatggaccc gccaccggtt ccgccaccat
     5401 atattcctga gttggcggcg atcgcagagg gaatcgggtt accgtctttc aaaagacctt
     5461 gcatggcctt gacaatcagc tcgcagtctg aattagtcgc aaagattggg atcgtaagct
     5521 ccatattaag ataagatcgc cacgcccctt ttggcacttc tacctttatg gtatattttc
     5581 ggttctgtgc ggaactttgg cgaactgagc atgtgacttt atatgcctga ctccggctgt
     5641 tactacttat ccattctgca atcccgttcg caaaattgct cggcgcaaca gttacgtctc
     5701 cggtgccacc attgtcaacc aacacaaatt gcgtaaagtt gctcgccata ccggtacctc
     5761 cccccgaacc gccactacca ccagaccccc ccgatccgcc gttgtcgtca tcgtccttgt
     5821 agtctcctcc agatccgcca gtgccacccc cagaccctcc tgtcccggcg cctctagtct
     5881 ttgagattcg tcggaacaca tgataataga ggtcccaggc ttggaggagg tccttgacat
     5941 tccctgattt catgtacttc ctgcaccact cttgggcctc cattaaatct cgaccatagg
     6001 cctgattaaa ggatgtttcc ttcagagtct ggggtccccg ttccatcata gcatgcaagg
     6061 gctccagcac ctcaaacatg cctttcacgt tcctttcccc aaagtacaaa cgagatgcct
     6121 cttccaggcc ttcatgccac atctccaggg atcccatggt ggcgaagctt aagtttaaac
     6181 gctagagtcc ggaggctgga tcggtcccgg tgtcttctat ggaggtcaaa acagcgtgga
     6241 tggcgtctcc aggcgatctg acggttcact aaacgagctc gtcgacgatc tctatcactg
     6301 atagggagat ctc
//
