LOCUS       sequence-487534-        6024 bp    DNA     circular SYN 30-SEP-2025
DEFINITION  Reporter for human p38alpha activation state by FRET.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6024)
  TITLE     pUC57-kan-VPC
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6024)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 30, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pUC-GW-Kan-VPC
FEATURES             Location/Qualifiers
     source          1..6024
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        476..779
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        780..983
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     934..954
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(1046..1048,1049..1051,1052..1762)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     RBS             2757..2765
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             join(2849..2851,2852..2854,2855..3568)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAIHGNVYITADKQKNGIK
                     ANFGLNCNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    3692..3813
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3729..3748)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3783..3802
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3848..3864)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3848..3864)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3861..3883)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3872..3888
                     /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(3896..3902,3903..3920,3921..3926))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3941..3962
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4079..4096)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4250..4838)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4330..4349)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5009..5824)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(5009..5824)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5732..5751
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(5825..5929)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5997..6015
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gacgcgtatt gggattagtt
      421 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
      481 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      541 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      601 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      661 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      721 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      781 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      841 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      901 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      961 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag cgcttaatac
     1021 gactcactat agaggaggtt gtacaatggt gagcaagggc gaggagctgt tcaccggggt
     1081 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     1141 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg
     1201 caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcc tgcagtgctt
     1261 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     1321 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     1381 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     1441 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     1501 tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat
     1561 cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     1621 ccccgtgctg ctgcccgaca accactacct gagctaccag tccaagctga gcaaagaccc
     1681 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     1741 cggcatggac gagctgtaca agggaatgtc tcaggagagg cccacgttct accggcagga
     1801 gctgaacaag acaatctggg aggtgcccga gcgttaccag aacctgtctc cagtgggctc
     1861 tggcgcctat ggctctgtgt gtgctgcttt tgacacaaaa acggggttac gtgtggcagt
     1921 gaagaagctc tccagaccat ttcagtccat cattcatgcg aaaagaacct acagagaact
     1981 gcggttactt aaacatatga aacatgaaaa tgtgattggt ctgttggacg tttttacacc
     2041 tgcaaggtct ctggaggaat tcaatgatgt gtatctggtg acccatctca tgggggcaga
     2101 tctgaacaac attgtgaaat gtcagaagct tacagatgac catgttcagt tccttatcta
     2161 ccaaattctc cgaggtctaa agtatataca ttcagctgac ataattcaca gggacctaaa
     2221 acctagtaat ctagctgtga atgaagactg tgagctgaag attctggatt ttggactggc
     2281 tcggcacaca gatgatgaaa tgacaggcta cgtggccact aggtggtaca gggctcctga
     2341 gatcatgctg aactggatgc attacaacca gacagttgat atttggtcag tgggatgcat
     2401 aatggccgag ctgttgactg gaagaacatt gtttcctggt acagaccata ttaaccagct
     2461 tcagcagatt atgcgtctga caggaacacc ccccgcttat ctcattaaca ggatgccaag
     2521 ccatgaggca agaaactata ttcagtcttt gactcagatg ccgaagatga actttgcgaa
     2581 tgtatttatt ggtgccaatc ccctggctgt cgacttgctg gagaagatgc ttgtattgga
     2641 ctcagataag agaattacag cggcccaagc ccttgcacat gcctactttg ctcagtacca
     2701 cgatcctgat gatgaaccag tggccgatcc ttatgatcag tcctttgaaa gcagggacct
     2761 ccttatagat gagtggaaaa gcctgaccta tgatgaagtc atcagctttg tgccaccacc
     2821 ccttgaccaa gaagagatgg agtccggaat ggtgagcaag ggcgaggagc tgttcaccgg
     2881 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     2941 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     3001 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgagctggg gcgtgcagtg
     3061 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     3121 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     3181 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     3241 caaggaggac ggcaacatcc tggggcacaa gctggagtac aacgccatcc acggcaacgt
     3301 ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcggcc tcaactgcaa
     3361 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     3421 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccaagc tgagcaaaga
     3481 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     3541 tctcggcatg gacgagctat acaagtaatt ataatctaga taactgatca taatcagcca
     3601 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     3661 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagcta ataatggtta
     3721 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3781 ttgtggtttg tccaaactca tcaatgtatc ttaatcacaa tggcgcgccg agcttggctc
     3841 gagcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     3901 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     3961 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4021 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4081 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4141 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4201 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4261 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4321 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4381 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4441 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4501 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4561 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4621 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4681 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4741 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4801 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4861 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4921 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4981 gtatatatga gtaaacttgg tctgacagtt agaaaaactc atcgagcatc aaatgaaact
     5041 gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg
     5101 aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga
     5161 ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat
     5221 caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca
     5281 tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat
     5341 caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt
     5401 taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat
     5461 caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccag
     5521 ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg
     5581 gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg
     5641 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc
     5701 gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat
     5761 cagcatccat gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc
     5821 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5881 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5941 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     6001 ggcgtatcac gaggcccttt cgtc
//
