LOCUS       sequence-421873-        7960 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Light-gated peroxidase (APEX) reporter for detecting 
            calcium-dependent CaM and MK2 interaction (component 2).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7960)
  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 7960)
  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 (TEVp)
FEATURES             Location/Qualifiers
     source          1..7960
                     /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             3058..3099
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3817..4405
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3870..3890)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4288..4299)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4477..4710
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4788..4909
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4825..4844)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4879..4898
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4949..5084
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5104..5123)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5105..5123)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5133..5150)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5133..5149)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5142..5164)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5291..5746
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5378..5397)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5588..5609
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5772..5876
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5877..5945,5946..6737)
                     /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(6095..6114)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6908..7496
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7397..7416
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7650..7667
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7784..7805
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7820..7825,7826..7843,7844..7850)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7858..7874
                     /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     7863..7885
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7882..7898
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7882..7898
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7917..7937
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7919..7937
                     /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 accatggacc aactgactga agagcagatc
     2641 gcagaattta aagaggcttt ctccctattt gacaaggacg gggatgggac aataacaacc
     2701 aaggagctgg ggacggtgat gcggtctctg gggcagaacc ccacagaagc agagctgcag
     2761 gacatgatca atgaagtaga tgccgacggt gacggcacaa tcgacttccc tgagttcctg
     2821 acaatgatgg caagaaaaat gaaatacagg gacacggaag aagaaattag agaagcgttc
     2881 cgtgtgtttg ataaggatgg caatggctac atcagtgcag cagagcttcg ccacgtgatg
     2941 acaaaccttg gagagaagtt aacagatgaa gaggttgatg aaatgatcag ggaagcagac
     3001 atcgatgggg atggtcaggt aaactacgaa gagtttgtac aaatgatgac agcgaagggc
     3061 aagccgatcc ctaaccctct gctgggcctg gacagcactg gtggagctag cggtggaagt
     3121 ggatcaggca gcggtggatc tggagaaagc ttgtttaagg gaccacgtga ttacaacccg
     3181 atatcgagca ccatttgtca tttgacgaat gaatctgatg ggcacacaac atcgttgtat
     3241 ggtattggat ttggtccctt catcattaca aacaagcact tgtttagaag aaataatgga
     3301 acactgttgg tccaatcact acatggtgta ttcaaggtca agaacaccac gactttgcaa
     3361 caacacctca ttgatgggag ggacatgata attattcgca tgcctaagga tttcccacca
     3421 tttcctcaaa agctgaaatt tagagagcca caaagggaag agcgcatatg tcttgtgaca
     3481 accaacttcc aaactaagag catgtctagc atggtgtcag acactagttg cacattccct
     3541 tcatctgatg gcatattctg gaagcattgg attcaaacca aggatgggca gtgtggcaat
     3601 ccattagtat caactagaga tgggttcatt gttggtatac actcagcatc gaatttcacc
     3661 aacacaaaca attatttcac aagcgtgccg aaaaacttca tggaattgtt gacaaatcag
     3721 gaggcgcagc agtgggttag tggttggcga ttaaatgctg actcagtatt gtggggtggc
     3781 cataaagttt tcatggtgta aacgcgttaa gtcgacaatc aacctctgga ttacaaaatt
     3841 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     3901 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     3961 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     4021 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     4081 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     4141 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     4201 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     4261 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     4321 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     4381 atctcccttt gggccgcctc cccgcgtcga ctttaagacc aatgacttac aaggcagctg
     4441 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     4501 gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     4561 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     4621 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     4681 ccttttagtc agtgtggaaa atctctagca gtacgtatag tagttcatgt catcttatta
     4741 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
     4801 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4861 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
     4921 tctagctatc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4981 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     5041 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat
     5101 tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac
     5161 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     5221 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     5281 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     5341 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     5401 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     5461 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     5521 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     5581 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     5641 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     5701 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     5761 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     5821 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     5881 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     5941 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     6001 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     6061 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     6121 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     6181 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     6241 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     6301 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     6361 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     6421 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     6481 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     6541 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     6601 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     6661 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     6721 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     6781 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6841 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6901 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6961 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     7021 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     7081 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7141 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7201 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7261 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7321 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7381 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7441 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7501 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     7561 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     7621 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     7681 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     7741 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     7801 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     7861 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa
     7921 ttaaccctca ctaaagggaa caaaagctgg agctgcaagc
//
