LOCUS       sequence-261550-        6628 bp    DNA     circular SYN 14-APR-2025
DEFINITION  GNCR1-BFP-CAAX.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6628)
  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 6628)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: GWF148
FEATURES             Location/Qualifiers
     source          1..6628
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    206..224
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    227..245
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     255..279
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      382..391
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1090..1119
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1132..1827
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     primer_bind     complement(1966..1983)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1972..2196
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2329..2348)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    2480..2527
                     /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             2535..3557
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             2535..3557
                     /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    3687..3808
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3724..3743)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3778..3797
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3857..3873)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3857..3873)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3870..3892)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3881..3897
                     /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(3905..3911,3912..3929,3930..3935))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3950..3971
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4088..4105)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4259..4847)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4339..4358)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5018..5809,5810..5878))
                     /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     5641..5660
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5879..5983)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6058..6077)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6249..6628
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctctccct atcagtgata gagatctccc tatcagtgat
      241 agagatcgtc gacgagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc
      301 tgttttgacc tccatagaag acaccgggac cgatccagcc tccggactct agcgtttaaa
      361 cttaagcttg gtaccgagct cgccaccatg gatatcgaga agctttgcaa aaaagctgaa
      421 gaagaggcaa aggaggcgca agagaaagct gatgagttgc ggcaacggca ccccgacagc
      481 caggctgctg aggatgctga ggacttggca aatctggcgg tggccgctgt cttgacggct
      541 tgcttgttgg cccaagaaca tccaaacgct gacattgcaa agctttgcat taaggccgcg
      601 tccgaagcgg ctgaagcagc ttctaaggca gccgagctgg cgcaacggca tcctgactcc
      661 caagcggcga gggacgctat aaaactcgca tcacaagcag ccagagctgt aatcctcgct
      721 attatgctcg cagccgaaaa cccaaacgct gatattgcca aactgtgtat aaaggcggct
      781 agtgaagccg cagaagctgc aagcaaagca gcagaacttg cccaacgcca ccccgactcc
      841 caggcggcta gagatgctat aaaacttgct tctcaggctg cggaggcggt agaacgagcg
      901 atctggctgg cagcagagaa ccccaacgct gacattgcca aaaagtgtat caaggcggct
      961 tccgaagccg ccgaggaggc cagcaaagcg gcggaagagg cacagagaca tccggactca
     1021 cagaaagcac gcgacgagat taaagaggct agtcaaaaag cagaggaggt aaaagaacgc
     1081 tgtaagtccg agcaaaaact catatctgag gaggacttgg gctcaggaag cagcgagctg
     1141 attaaggaga acatgcacat gaagctgtac atggagggca ccgtggacaa ccatcacttc
     1201 aagtgcacat ccgagggcga aggcaagccc tacgagggca cccagaccat gagaatcaag
     1261 gtggtcgagg gcggccctct ccccttcgcc ttcgacatcc tggctactag cttcctctac
     1321 ggcagcaaga ccttcatcaa ccacacccag ggcatccccg acttcttcaa gcagtccttc
     1381 cctgagggct tcacatggga gagagtcacc acatacgaag acgggggcgt gctgaccgct
     1441 acccaggaca ccagcctcca ggacggctgc ctcatctaca acgtcaagat cagaggggtg
     1501 aacttcacat ccaacggccc tgtgatgcag aagaaaacac tcggctggga ggccttcacc
     1561 gagacgctgt accccgctga cggcggcctg gaaggcagaa acgacatggc cctgaagctc
     1621 gtgggcggga gccatctgat cgcaaacatc aagaccacat atagatccaa gaaacccgct
     1681 aagaacctca agatgcctgg cgtctactat gtggactaca gactggaaag aatcaaggag
     1741 gccaacaacg agacctacgt cgagcagcac gaggtggcag tggccagata ctgcgacctc
     1801 cctagcaaac tggggcacaa gcttaatcga aaacataaag aaaagatgag caaagatggt
     1861 aaaaagaaga aaaagaagtc aaagacaaag tgtgtaatta tgtgataaat ccagcacagt
     1921 ggcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt
     1981 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     2041 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     2101 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     2161 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     2221 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     2281 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     2341 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     2401 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     2461 gtgatggttc acgtacctag aagttcctat tccgaagttc ctattctcta gaaagtatag
     2521 gaacttcctt ggccaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg
     2581 aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt
     2641 tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt
     2701 tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg attccggaag
     2761 tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc cgtgcacagg
     2821 gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg
     2881 aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc ggcccattcg
     2941 gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc
     3001 cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg
     3061 ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg
     3121 cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact
     3181 ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc
     3241 cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg
     3301 caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga
     3361 gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg
     3421 tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct
     3481 ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc
     3541 cgagggcaaa ggaatagcac gtactacgag atttcgattc caccgccgcc ttctatgaaa
     3601 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     3661 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     3721 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3781 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     3841 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     3901 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     3961 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4021 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4081 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4141 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4201 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4261 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4321 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4381 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4441 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4501 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4561 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4621 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4681 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4741 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4801 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4861 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4921 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4981 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5041 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5101 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5161 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5221 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5281 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5341 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5401 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5461 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5521 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5581 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5641 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5701 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5761 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5821 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5881 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5941 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6001 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
     6061 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     6121 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
     6181 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
     6241 tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     6301 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     6361 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     6421 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     6481 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     6541 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     6601 ctacgtatta gtcatcgcta ttaccatg
//
