LOCUS       sequence-421894-        8788 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Peroxidase (APEX) reporter for beta 2-adrenergic receptor (beta-2AR)
            and beta-arrestin 2 interaction (component 2).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8788)
  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 8788)
  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: beta-2AR-PPI-CALPon1.0 (TEVp)
FEATURES             Location/Qualifiers
     source          1..8788
                     /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             2617..2643
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    2650..2674
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3902..3926)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    4645..5233
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4698..4718)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5116..5127)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5305..5538
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5616..5737
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5653..5672)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5707..5726
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5777..5912
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5932..5951)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5933..5951)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5961..5978)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5961..5977)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5970..5992)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6119..6574
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6206..6225)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6416..6437
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6600..6704
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6705..6773,6774..7565)
                     /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(6923..6942)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7736..8324
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8225..8244
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8478..8495
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8612..8633
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8648..8653,8654..8671,8672..8678)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8686..8702
                     /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     8691..8713
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8710..8726
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8710..8726
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8745..8765
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8747..8765
                     /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 accatgtacc catatgacgt tccagattac
     2641 gctaccggtc caactttgta caaaaaagtt ggcatgggtg aaaaacccgg gaccagggtc
     2701 ttcaagaagt cgagccctaa ctgcaagctc accgtgtact tgggcaagcg tgactttgtg
     2761 gatcacttgg acaaagtgga tcctgtcgat ggtgtggtgc ttgtggatcc tgactacttg
     2821 aaggaccgga aagtgtttgt gaccctcacc tgtgccttcc gctatggccg agaagacctg
     2881 gatgtactgg gcctgtcttt ccgcaaagat ctgttcatcg ccacctacca ggccttcccc
     2941 cccatgccca acccacctcg gccccccacc cgcctacagg accgactgct gaagaagttg
     3001 ggccagcacg cccacccctt ttttttcaca ataccccaga atttgccttg ctccgtcaca
     3061 ctgcagccag gaccggagga cacagggaag gcctgtggag tagactttga gattcgagcc
     3121 ttctgtgcca aatctataga agaaaaaagc cacaaaagga actccgtgcg gcttatcatc
     3181 agaaaggtac agtttgctcc tgagacaccc ggcccccagc catcagctga aaccacacgc
     3241 cacttcctca tgtctgaccg gaggtccctg cacctagagg cttccctgga caaagagctg
     3301 tactaccatg gggaacccct caatgtcaac gtccacgtca ccaacaattc tgccaaaacc
     3361 gtcaagaaga tcagagtgtc tgtgagacag tatgccgaca tttgcctctt cagcaccgcg
     3421 cagtacaagt gtcctgtggc tcagcttgaa caagatgacc aggtgtctcc cagttccaca
     3481 ttctgcaagg tgtacaccat aaccccgctg ctcagtgaca accgagagaa gcgtggcctt
     3541 gcccttgatg ggcaactcaa gcacgaagac accaacctgg cttccagcac cattgtgaag
     3601 gagggagcca acaaggaggt gctgggaatc ctagtatcct acagggtcaa ggtgaagctg
     3661 gtggtgtctc gaggcgggga tgtctccgtg gagctacctt tcgtcctaat gcaccccaag
     3721 ccccacgacc acatcaccct tccccgaccc cagtcagccc cccgggaaat agacatccct
     3781 gtggatacca acctcattga attcgatacc aactatgcca cagacgacga catcgtgttt
     3841 gaggactttg cgaggcttcg gctgaagggg atgaaggatg acgactgtga tgaccagttc
     3901 tgcccaactt tcttgtacaa agttggcatt atagctagcg gtggaagtgg atcaggcagc
     3961 ggtggatctg gagaaagctt gtttaaggga ccacgtgatt acaacccgat atcgagcacc
     4021 atttgtcatt tgacgaatga atctgatggg cacacaacat cgttgtatgg tattggattt
     4081 ggtcccttca tcattacaaa caagcacttg tttagaagaa ataatggaac actgttggtc
     4141 caatcactac atggtgtatt caaggtcaag aacaccacga ctttgcaaca acacctcatt
     4201 gatgggaggg acatgataat tattcgcatg cctaaggatt tcccaccatt tcctcaaaag
     4261 ctgaaattta gagagccaca aagggaagag cgcatatgtc ttgtgacaac caacttccaa
     4321 actaagagca tgtctagcat ggtgtcagac actagttgca cattcccttc atctgatggc
     4381 atattctgga agcattggat tcaaaccaag gatgggcagt gtggcaatcc attagtatca
     4441 actagagatg ggttcattgt tggtatacac tcagcatcga atttcaccaa cacaaacaat
     4501 tatttcacaa gcgtgccgaa aaacttcatg gaattgttga caaatcagga ggcgcagcag
     4561 tgggttagtg gttggcgatt aaatgctgac tcagtattgt ggggtggcca taaagttttc
     4621 atggtgtaaa cgcgttaagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     4681 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     4741 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     4801 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     4861 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     4921 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     4981 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     5041 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     5101 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     5161 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     5221 gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     5281 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     5341 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     5401 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     5461 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     5521 tgtggaaaat ctctagcagt acgtatagta gttcatgtca tcttattatt cagtatttat
     5581 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
     5641 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5701 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
     5761 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     5821 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     5881 ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt
     5941 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     6001 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     6061 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     6121 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     6181 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     6241 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     6301 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     6361 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     6421 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     6481 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     6541 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     6601 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     6661 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     6721 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     6781 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     6841 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     6901 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     6961 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     7021 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     7081 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     7141 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     7201 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     7261 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     7321 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     7381 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     7441 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     7501 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     7561 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     7621 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     7681 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     7741 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     7801 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     7861 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     7921 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     7981 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     8041 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     8101 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     8161 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     8221 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     8281 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     8341 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     8401 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     8461 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     8521 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     8581 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     8641 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     8701 aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact
     8761 aaagggaaca aaagctggag ctgcaagc
//
