LOCUS       sequence-261551-        6502 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NS3aH1-mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6502)
  AUTHORS   Foight GW, Wang Z, Wei CT, Jr Greisen P, Warner KM, 
            Cunningham-Bryant D, Park K, Brunette TJ, Sheffler W, Baker D, Maly 
            DJ
  TITLE     Multi-input chemical control of protein dimerization for programming
            graded cellular responses.
  JOURNAL   Nat Biotechnol. 2019 Sep 9. pii: 10.1038/s41587-019-0242-8. doi: 
            10.1038/s41587-019-0242-8.
  PUBMED    31501561
REFERENCE   2  (bases 1 to 6502)
  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 
            Biotechnol. 2019 Sep 9. pii: 10.1038/s41587-019-0242-8. doi: 
            10.1038/s41587-019-0242-8."
COMMENT     Sequence Label: GWF094
FEATURES             Location/Qualifiers
     source          1..6502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             625..1332
                     /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(770..788)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1038..1057
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1348..1371
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1488..1505)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1494..1718
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1851..1870)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    2002..2049
                     /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)."
     CDS             2057..3079
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             2057..3079
                     /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
                     "
     polyA_signal    3209..3330
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3246..3265)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3300..3319
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3379..3395)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3379..3395)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3392..3414)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3403..3419
                     /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)."
     promoter        complement(join(3427..3433,3434..3451,3452..3457))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3472..3493
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3610..3627)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3781..4369)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3861..3880)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4540..5331,5332..5400))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRHIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5163..5182
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5401..5505)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5580..5599)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5771..6150
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6151..6354
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6305..6325
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    6356..6374
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6377..6395
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     6405..6429
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 cgtttaaaca tgatgaaaaa gaaaggttct gttgtcatcg ttggacgcat aaatctctct
       61 ggcgacaccg catattctca gcagacgcga ggtctggaag ggtgtcaaga gacctcacaa
      121 accgggaggg acaagaatca agtagaaggg gaggtccagg tggtatctac ggcaacccaa
      181 tccttcttgg cgacatctat caatggagtc ctgtggacgg tttatcatgg tgcgggcact
      241 cggacaattg cctctccaaa aggtccagtt acccagatgt atactaatgt agacaaagat
      301 cttgtcggat ggcaagcacc acaagggtct cggagcctca ctccctgtac gtgtggaagt
      361 tcagaccttt atctcgtaac tagacacgcg gatgttatcc ccgttaggag aagaggagat
      421 agtaggggat ccctgctttc accacgccca atatcctatc ttaagggttc ttcaggtggt
      481 cccttgttgt gtccggcggg tcacgcggta ggtatattcc gcgcggccgt atctactcgg
      541 ggcgtggcta aggcggtcga tttcataccg gttgagtcct tggaaacaac gatgcggagc
      601 cccggttcag ggacaggctc tggtatggtg agcaagggcg aggaggataa catggccatc
      661 atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc
      721 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
      781 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac
      841 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc
      901 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
      961 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc
     1021 aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc
     1081 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg
     1141 aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg
     1201 cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac
     1261 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
     1321 gagctgtaca agggtagcgg aactggggac tacaaggacg acgacgacaa gtgattaagc
     1381 ttggtaccga gctcggatcc actagtccag tgtggtggaa ttctgcagat atccagcaca
     1441 gtggcggccg ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     1501 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     1561 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     1621 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     1681 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     1741 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     1801 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     1861 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     1921 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     1981 gggtgatggt tcacgtacct agaagttcct attccgaagt tcctattctc tagaaagtat
     2041 aggaacttcc ttggccaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat
     2101 cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc
     2161 tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg
     2221 tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga
     2281 agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca
     2341 gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc
     2401 ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt
     2461 cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga
     2521 tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca
     2581 ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca
     2641 cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga
     2701 ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag
     2761 gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct
     2821 tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca
     2881 gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat
     2941 cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt
     3001 ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcacccg
     3061 tccgagggca aaggaatagc acgtactacg agatttcgat tccaccgccg ccttctatga
     3121 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     3181 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     3241 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3301 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     3361 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     3421 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     3481 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     3541 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3601 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3661 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3721 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3781 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3841 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3901 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3961 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4021 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4081 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4141 taacaggatt agcaaagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4201 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4261 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4321 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4381 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4441 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4501 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4561 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4621 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4681 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4741 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4801 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4861 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4921 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4981 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5041 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5101 caagtcattc tgagaatagt gtatgtggcg accgagttgc tcttgcccgg cgtcaatacg
     5161 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5221 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5281 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5341 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5401 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5461 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5521 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     5581 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     5641 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     5701 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
     5761 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     5821 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     5881 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     5941 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     6001 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     6061 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     6121 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     6181 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     6241 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgaccca
     6301 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctccct
     6361 atcagtgata gagatctccc tatcagtgat agagatcgtc gacgagctcg tttagtgaac
     6421 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac
     6481 cgatccagcc tccggactct ag
//
