LOCUS       sequence-423195-        8593 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the CapC-caged opioid peptide in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8593)
  AUTHORS   Shen J, Geng L, Li X, Emery C, Kroning K, Shingles G, Lee K, Heyden 
            M, Li P, Wang W
  TITLE     A general method for chemogenetic control of peptide function.
  JOURNAL   Nat Methods. 2023 Jan;20(1):112-122. doi: 
            10.1038/s41592-022-01697-8. Epub 2022 Dec 8.
  PUBMED    36481965
REFERENCE   2  (bases 1 to 8593)
  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: 
            "10.1038/s41592-022-01697-8"; journalName: "Nat Methods"; date: 
            "2023-01"; volume: "20"; issue: "1"; pages: "112-122"
COMMENT     Sequence Label: plx208-CMV-Signal
            seuqence-enkephalin-CapC-FLA...
FEATURES             Location/Qualifiers
     source          1..8593
                     /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             2737..3057
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=FKBP-V
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             3109..3132
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    4450..5038
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4503..4523)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4921..4932)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5110..5343
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5421..5542
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5458..5477)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5512..5531
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5582..5717
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5737..5756)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5738..5756)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5766..5783)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5766..5782)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5775..5797)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5924..6379
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6011..6030)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6221..6242
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6405..6509
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6510..6578,6579..7370)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIVAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6728..6747)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7541..8129
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8030..8049
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8283..8300
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8417..8438
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8453..8458,8459..8476,8477..8483)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8491..8507
                     /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     8496..8518
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8515..8531
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8515..8531
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8550..8570
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8552..8570
                     /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 accatgaaga ccatcatcgc cctgagctac
     2641 atcttctgcc tggtgttcgc ctacggcgga tttatgccta atttgcggcc ttttggttca
     2701 ggcggttctg gtactggatc tggttctgga ggttctggag tgcaggtgga aaccatctcc
     2761 ccaggagacg ggcgcacctt ccccaagcgc ggccagacct gcgtggtgca ctacaccggg
     2821 atgcttgaag atggaaagaa agttgattcc tcccgggaca gaaacaagcc ctttaagttt
     2881 atgctaggca agcaggaggt gatccgaggc tgggaagaag gggttgccca gatgagtgtg
     2941 ggtcagagag ccaaactgac tatatctcca gattatgcct atggtgccac tgggcaccca
     3001 ggcatcatcc caccacatgc cactctcgtc ttcgatgtgg agcttctaaa actggaaacc
     3061 ggtggtggaa gtggatcagg cagcggtgga tctggaggta gcggcggaga ctacaaggat
     3121 gacgacgata agggcggaat ggactcttcc gctgctccca ccaatgcctc aaattgtaca
     3181 gacgccttgg cctatagctc ttgctctccc gcaccatctc cgggaagctg ggttaacctc
     3241 agtcatctcg acgggaacct gtccgaccct tgtggcccta ataggaccga cctcggtggt
     3301 cgagatagcc tgtgcccccc tactggaagc cccagcatga tcacagctat cacaataatg
     3361 gctctctata gtatcgtctg cgtcgtggga ctctttggca attttctcgt catgtacgtt
     3421 atcgtcagat acacaaagat gaaaaccgca accaatattt acatctttaa tctggccctg
     3481 gcggatgccc ttgcaaccag tacactgccg tttcaatccg tgaattatct gatgggcaca
     3541 tggcccttcg gaaccattct gtgcaaaatc gtaatctcaa ttgattatta taacatgttc
     3601 acctccattt ttaccttgtg tacaatgtca gtagaccgct atatcgccgt atgccaccct
     3661 gtcaaagctc ttgatttcag gacccctagg aatgccaaga tcatcaatgt gtgcaattgg
     3721 attctgtcta gcgctatcgg cttgccggtg atgttcatgg ccactaccaa gtacagacaa
     3781 ggttcaatcg actgtaccct cacattttca catcccacgt ggtattggga gaacttgctg
     3841 aagatctgtg tgttcatctt cgccttcatc atgccagtac tgataatcac cgtttgttat
     3901 ggtcttatga tcctgcgcct gaagagcgtg cgcatgctca gcggctccaa agaaaaggac
     3961 cggaaccttc ggcgaatcac acggatggtc ctggtggttg tggctgtgtt cattgtttgc
     4021 tggaccccta ttcatattta tgttataatt aaggccctgg tgaccatccc ggaaaccaca
     4081 ttccagaccg tttcctggca cttttgtata gcattgggat ataccaacag ttgccttaat
     4141 cccgttctgt atgcttttct ggacgagaat ttcaagcgat gcttcaggga gttctgcatt
     4201 cccacaagct ctaacattga acagcagaat tctacacgaa tacgacagaa taccagagat
     4261 catcctagca ctgccaatac tgtcgaccgg accaatcacc agcttgagaa ccttgaggca
     4321 gaaaccgccc ctctcccaga gctggccgag aagctggccg gcctggacat caacggtgga
     4381 gctagcggaa gcggctcggt gacgggttat agattgttcg aggagattct gtagacgcgt
     4441 taagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     4501 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     4561 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     4621 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     4681 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     4741 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     4801 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     4861 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     4921 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     4981 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcgt
     5041 cgactttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     5101 aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta
     5161 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     5221 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     5281 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     5341 gcagtacgta tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat
     5401 gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca
     5461 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     5521 ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc
     5581 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     5641 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     5701 aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc gtattacgcg
     5761 cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     5821 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     5881 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc
     5941 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     6001 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     6061 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     6121 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     6181 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     6241 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     6301 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     6361 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     6421 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     6481 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     6541 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     6601 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     6661 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     6721 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     6781 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     6841 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6901 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     6961 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     7021 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     7081 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     7141 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     7201 ataaatctgg agccggtgag cgtgggtctc gcggtatcgt tgcagcactg gggccagatg
     7261 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     7321 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     7381 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     7441 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     7501 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     7561 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     7621 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     7681 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     7741 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     7801 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     7861 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     7921 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     7981 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     8041 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     8101 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     8161 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     8221 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     8281 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     8341 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     8401 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     8461 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     8521 acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc
     8581 tggagctgca agc
//
