LOCUS       sequence-365868-        5335 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cyclofen-inducible PK inhibitor alpha (mut) activation via mammalian
            cell transfection or mRNA synthesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5335)
  TITLE     Vriz lab cyclofen-inducible ERT2 plasmids
REFERENCE   2  (bases 1 to 5335)
  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"
COMMENT     Sequence Label: pCSZPKIamutHAERT(#119)
FEATURES             Location/Qualifiers
     source          1..5335
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             337..363
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             421..1356
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             469..1362
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATA"
     polyA_signal    1387..1521
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1411..1430)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1465..1484
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1634..1654)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1634..1652)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1673..1689)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1673..1689)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1686..1708)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1697..1713
                     /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(1721..1727,1728..1745,1746..1751))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1766..1787
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1904..1921)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2075..2663)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2155..2174)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2834..3625,3626..3694))
                     /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     3457..3476
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3695..3799)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3825..4280)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3962..3983)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4174..4193
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4351..5335
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccgctagc accggtgagc tcaccatgac tgatgtggaa
      121 cctgttgtct ctgactttgc agccacgggc aggactggtg cagcgaatgc cttgccagac
      181 attctaggtt ccacggctgg tcctggagcc gccgacctac ctgataaact agccgaactc
      241 acagtaggag atgatggtga acagggtgga gagagttcat cctcaaccga cccacccaaa
      301 gaatccacag agggggcaaa gacagagggt tcagtatacc catacgatgt tccagattac
      361 gcccacgcgt ccggtccgag cgcttccgga cctaggccgg gtcccgggct cgagccatct
      421 gctggagaca tgagagctgc caacctttgg ccaagcccgc tcatgatcaa acgctctaag
      481 aagaacagcc tggccttgtc cctgacggcc gaccagatgg tcagtgcctt gttggatgct
      541 gagcccccca tactctattc cgagtatgat cctaccagac ccttcagtga agcttcgatg
      601 atgggcttac tgaccaacct ggcagacagg gagctggttc acatgatcaa ctgggcgaag
      661 agggtgccag gctttgtgga tttgaccctc catgatcagg tccaccttct agaatgtgcc
      721 tggctagaga tcctgatgat tggtctcgtc tggcgctcca tggagcaccc agtgaagcta
      781 ctgtttgctc ctaacttgct cttggacagg aaccagggaa aatgtgtaga gggcatggtg
      841 gagatcttcg acatgctgct ggctacatca tctcggttcc gcatgatgaa tctgcaggga
      901 gaggagtttg tgtgcctcaa atctattatt ttgcttaatt ctggagtgta cacatttctg
      961 tccagcaccc tgaagtctct ggaagagaag gaccatatcc accgagtcct ggacaagatc
     1021 acagacactt tgatccacct gatggccaag gcaggcctga ccctgcagca gcagcaccag
     1081 cggctggccc agctcctcct catcctctcc cacatcaggc acatgagtaa caaaggcatg
     1141 gagcatctgt acagcatgaa gtgcaagaac gtggtgcccc tctatgacct gctgctggag
     1201 gcggcggacg cccaccgcct acatgcgccc actagccgtg gaggggcatc cgtggaggag
     1261 acggaccaaa gccacttggc cactgcgggc tctacttcat cgcattcctt gcaaaagtat
     1321 tacatcacgg gggaggcaga gggtttccct gccacagctt gatgaagatc tgagggcccg
     1381 tttgtagatc cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     1441 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     1501 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     1561 ggggaggtgt gggaggtttt ttaattcgcg gccgcggcgc caatgcattg ggcccggtac
     1621 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
     1681 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccgggagca
     1741 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     1801 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     1861 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     1921 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     1981 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2041 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2101 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2161 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2221 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2281 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2341 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2401 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     2461 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     2521 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     2581 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     2641 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     2701 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     2761 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     2821 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     2881 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     2941 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3001 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3061 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3121 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     3181 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     3241 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     3301 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     3361 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     3421 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     3481 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     3541 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     3601 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     3661 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     3721 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     3781 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa
     3841 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     3901 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     3961 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     4021 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     4081 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     4141 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     4201 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     4261 tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg
     4321 atcggtgcgg gcctcttcgc tattacgcca gtcgaccgcc aattcaatat ggcgtatatg
     4381 gactcatgcc aattcaatat ggtggatctg gacctgtgcc aattcaatat ggcgtatatg
     4441 gactcgtgcc aattcaatat ggtggatctg gaccccagcc aattcaatat ggcggacttg
     4501 gcaccatgcc aattcaatat ggcggacctg gcactgtgcc aactggggag gggtctactt
     4561 ggcacggtgc caagtttgag gaggggtctt ggccctgtgc caagtccgcc atattgaatt
     4621 ggcatggtgc caataatggc ggccatattg gctatatgcc aggatcaata tataggcaat
     4681 atccaatatg gccctatgcc aatatggcta ttggccaggt tcaatactat gtattggccc
     4741 tatgccatat agtattccat atatgggttt tcctattgac gtagatagcc cctcccaatg
     4801 ggcggtccca tataccatat atggggcttc ctaataccgc ccatagccac tcccccattg
     4861 acgtcaatgg tctctatata tggtctttcc tattgacgtc atatgggcgg tcctattgac
     4921 gtatatggcg cctcccccat tgacgtcaat tacggtaaat ggcccgcctg gctcaatgcc
     4981 cattgacgtc aataggacca cccaccattg acgtcaatgg gatggctcat tgcccattca
     5041 tatccgttct cacgccccct attgacgtca atgacggtaa atggcccact tggcagtaca
     5101 tcaatatcta ttaatagtaa cttggcaagt acattactat tggaagtacg ccagggtaca
     5161 ttggcagtac tcccattgac gtcaatggcg gtaaatggcc cgcgatggct gccaagtaca
     5221 tccccattga cgtcaatggg gaggggcaat gacgcaaatg ggcgttccat tgacgtaaat
     5281 gggcggtagg cgtgcctaat gggaggtcta tataagcaat gctcgtttag ggaac
//
