LOCUS       sequence-421876-        8443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Light-gated peroxidase (APEX) reporter for detecting 
            calcium-dependent CaM and MK2 interaction (component 1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8443)
  AUTHORS   Zhou G, Wan WW, Wang W
  TITLE     Modular Peroxidase-Based Reporters for Detecting Protease Activity 
            and Protein Interactions with Temporal Gating.
  JOURNAL   J Am Chem Soc. 2022 Dec 21;144(50):22933-22940. doi: 
            10.1021/jacs.2c08280. Epub 2022 Dec 13.
  PUBMED    36511757
REFERENCE   2  (bases 1 to 8443)
  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"; doi: 
            "10.1021/jacs.2c08280"; journalName: "J Am Chem Soc"; date: 
            "2022-12-21- 21"; volume: "144"; issue: "50"; pages: "22933-22940"
COMMENT     Sequence Label: Ca-LiPPI-CLAPon1.0 (APEX)
FEATURES             Location/Qualifiers
     source          1..8443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..231
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             232..412
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    459..584
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1077..1310
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1495..1539
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1847..1964
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2081..2384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2385..2588
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2539..2559
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3322..3342
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQA"
     CDS             3610..3636
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3637..3663
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    4300..4888
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4353..4373)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4771..4782)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4960..5193
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5271..5392
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5308..5327)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5362..5381
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5432..5567
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5587..5606)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5588..5606)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5616..5633)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5616..5632)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5625..5647)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5774..6229
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5861..5880)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6071..6092
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6255..6359
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6360..6428,6429..7220)
                     /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(6578..6597)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7391..7979
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7880..7899
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8133..8150
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8267..8288
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8303..8308,8309..8326,8327..8333)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8341..8357
                     /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     8346..8368
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8365..8381
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8365..8381
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8400..8420
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8402..8420
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
       61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
      121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
      181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
      241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
      421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
      481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
      541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
      601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
      661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
      721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
      781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
      841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
      901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
      961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     1681 tttcagaccc acctcccaac cccgagagcg ctgccgtctg gaccgatggc tgtgtggaag
     1741 tgctcgccga tagtggaaac agacgcccca gcactcgtcc tagggcaaag gatctgcagt
     1801 aatgagaatt ctagatcttg agacaaatgg cagtattcat ccacaatttt aaaagaaaag
     1861 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     1921 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     1981 acagcagaga tccactttgg cgccggctcg agggggcccg gggttattaa tagtaatcaa
     2041 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     2101 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2161 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2221 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2281 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2341 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2401 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2461 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2521 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2581 gcagagctct ctggctaact gtcgggatcc accatgttca acgctcgcag gaagctggct
     2641 ggtgctatcc tgactaccat gctcgccaca agaaacttca gcggcagctt taatgcgcgg
     2701 cgcaagctag ccggagccat acttacgaca atgttggcta ccagaaattt ttcggagctg
     2761 gccgagaagc tggccggcct ggacatcaac ggtggagcta gcatggaagt cagcgccttg
     2821 gagaaggagg tctctgccct tgaaaaagag gtttcagccc tcgagaaaga agtctctgcg
     2881 ctcgaaaaag aagtgtcagc gcttgagaag cgggcaacca cactggaacg gatcgagaaa
     2941 agcttcgtga ttactgatcc gagactgcct gacaacccaa tcatttttgt gagcgattcc
     3001 ttcctgcagc tgacagaata ttctcgggaa gagatcctgg ggcgcaattg ccgttttctg
     3061 cagggacccg agacagaccg tgccactgtt cggaaaatca gagatgctat tgacaaccag
     3121 actgaagtga ccgttcagct gatcaattat accaagagcg gcaagaagtt ctggaacgtg
     3181 ttccacctgc agccgatgcg cgattataag ggcgacgtcc agtacttcat tggcgtgcag
     3241 ctggatggca ccgaacgtct tcatggcgcc gctgagcgtg aggcggtctg cctggtcaaa
     3301 aagacagcct ttcagattgc tgagaacctg tacttccagg cgaccggtgg gctgctgcag
     3361 ctgcccagcg acaaagccct gctgtccgat cccgtgttca gacctctggt cgataagtat
     3421 gcagccgacg aggatgcttt tttcgcagat tacgcagaag cacatcagaa gctgtcagaa
     3481 ctgggatttg ccgacgccgc tagcaaagtt tccgccctca aggaaaaggt cagtgctctt
     3541 aaagagaaag tatccgcact caaagagaag gttagcgcac tgaaagaaaa ggtcagcgca
     3601 ctgaaagaat acccatatga cgttccagat tacgcttacc catatgacgt tccagattac
     3661 gctagctcta gtggatcgac tagtgggaaa tcatacccaa cagtgtccgc agactaccag
     3721 gatgccgtgg agaaagccaa gaagaggctg ggagggttta tcgcagaaaa gaggtgcgca
     3781 cctctgatgc tgagactggc tttccacagc gcaggcacct ttgacaagcg aaccaaaaca
     3841 ggcggaccct ttggaacaat ccggcgccct gctgaactgg cacatagtgc caacagtggg
     3901 ctggacatcg ccgtgcggct gctggaacct ctgaaagcag agttcccaat tctgtcctac
     3961 gccgattttt atcagctggc aggagtggtc gctgtggagg tcactggggg ccccaaggtg
     4021 cctttccacc caggacggga ggacaaacca gaaccacctc cagaggggcg cctgccagat
     4081 ccgacaaagg gctccgacca tctgcgagat gtgtttggga aagctatggg cctgactgac
     4141 caggatatcg tcgcactgtc tggagggcac acccttggcg ccgctcataa ggaaaggtca
     4201 ggcttcgagg gaccctggac aagcaacccc ctgattttcg acaattctta ctttactgaa
     4261 ctgctgagtg gagagaagga ataaacgcgt taagtcgaca atcaacctct ggattacaaa
     4321 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     4381 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     4441 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     4501 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     4561 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     4621 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     4681 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
     4741 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     4801 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     4861 cggatctccc tttgggccgc ctccccgcgt cgactttaag accaatgact tacaaggcag
     4921 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     4981 aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag
     5041 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     5101 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     5161 gaccctttta gtcagtgtgg aaaatctcta gcagtacgta tagtagttca tgtcatctta
     5221 ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta
     5281 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5341 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5401 ggctctagct atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     5461 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     5521 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc
     5581 aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt
     5641 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     5701 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     5761 aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     5821 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     5881 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     5941 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     6001 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     6061 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     6121 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     6181 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
     6241 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     6301 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     6361 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     6421 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     6481 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     6541 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     6601 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     6661 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     6721 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     6781 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     6841 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     6901 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     6961 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     7021 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     7081 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     7141 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     7201 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     7261 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     7321 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     7381 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     7441 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     7501 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     7561 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     7621 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     7681 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     7741 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     7801 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     7861 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     7921 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     7981 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     8041 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     8101 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     8161 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     8221 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     8281 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     8341 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
     8401 caattaaccc tcactaaagg gaacaaaagc tggagctgca agc
//
