LOCUS       sequence-377950-        8245 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rapamycin-dependent opioid sensor for opioid receptors (component 
            2).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8245)
  AUTHORS   Kroning KE, Li M, Shen J, Fiel H, Nassar M, Wang W
  TITLE     A Modular Fluorescent Sensor Motif Used to Detect Opioids, 
            Protein-Protein Interactions, and Protease Activity.
  JOURNAL   ACS Chem Biol. 2022 Aug 19;17(8):2212-2220. doi: 
            10.1021/acschembio.2c00364. Epub 2022 Aug 4.
  PUBMED    35925780
REFERENCE   2  (bases 1 to 8245)
  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/acschembio.2c00364"; journalName: "ACS Chem Biol"; date: 
            "2022-08-19- 19"; volume: "17"; issue: "8"; pages: "2212-2220"
COMMENT     Sequence Label: Lentiviral-FKBP-cpGFP-Nb39
FEATURES             Location/Qualifiers
     source          1..8245
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..227
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             228..408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    455..580
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1073..1306
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1491..1535
                     /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    1843..1960
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2077..2380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2381..2584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2535..2555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(2610..2612,2613..2933)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     primer_bind     3176..3197
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3303..3324)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3564..3583)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4098..4686
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4151..4171)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4569..4580)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4758..4991
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5069..5190
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5106..5125)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5160..5179
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5230..5365
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5385..5404)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5386..5404)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5414..5431)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5414..5430)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5423..5445)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5572..6027
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5659..5678)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5869..5890
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6053..6157
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6158..6226,6227..7018)
                     /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(6376..6395)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7189..7777
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7678..7697
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7931..7948
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8065..8086
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8101..8106,8107..8124,8125..8131)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8139..8155
                     /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     8144..8166
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8163..8179
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8163..8179
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8198..8218
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8200..8218
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
       61 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
      121 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
      181 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt
      241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
      421 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
      481 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
      541 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
      601 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
      661 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
      721 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
      781 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
      841 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
      901 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
      961 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1021 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1081 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1141 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1201 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1261 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1321 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     1381 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     1441 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     1501 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     1561 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     1621 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     1681 agacccacct cccaaccccg agagcgctgc cgtctggacc gatggctgtg tggaagtgct
     1741 cgccgatagt ggaaacagac gccccagcac tcgtcctagg gcaaaggatc tgcagtaatg
     1801 agaattctag atcttgagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     1861 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     1921 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     1981 cagagatcca ctttggcgcc ggctcgaggg ggcccggggt tattaatagt aatcaattac
     2041 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     2101 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     2161 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     2221 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     2281 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     2341 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     2401 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     2461 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     2521 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     2581 agctctctgg ctaactgtcg ggatccacca tgggagtgca ggtggaaacc atctccccag
     2641 gagacgggcg caccttcccc aagcgcggcc agacctgcgt ggtgcactac accgggatgc
     2701 ttgaagatgg aaagaaattt gattcctccc gggacagaaa caagcccttt aagtttatgc
     2761 taggcaagca ggaggtgatc cgaggctggg aagaaggggt tgcccagatg agtgtgggtc
     2821 agagagccaa actgactata tctccagatt atgcctatgg tgccactggg cacccaggca
     2881 tcatcccacc acatgccact ctcgtcttcg atgtggagct tctaaaactg gaaggcggca
     2941 gtggcagtag tggcggcagt ggctctagaa acgtctatat caaggccgac aagcagaaga
     3001 acggcatcaa ggcgaacttc cacatccgcc acaacatcga ggacggcggc gtgcagctcg
     3061 cctaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     3121 actacctgag cgtgcagtcc aaactttcga aagaccccaa cgagaagcgc gatcacatgg
     3181 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg
     3241 gcggtaccgg agggagcatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     3301 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggtg
     3361 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     3421 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     3481 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacatcc
     3541 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     3601 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     3661 gcaacatcct ggggcacaag ctggagtaca accttaagga agatatcatg gctcaagtcc
     3721 aacttgtaga gtctggagga ggtctcgtgc ggcccggcgg tagcctgcga ctctcctgtg
     3781 ttgattcaga gaggacgagc taccctatgg ggtggtttag acgggcaccc ggaaaggagc
     3841 gcgaattcgt tgcaagtatc acgtggagtg gcattgaccc gacttatgct gatagcgttg
     3901 ccgaccggtt tactacgtcc agagacgttg cgaataacac gctgtacctc cagatgaact
     3961 ctctcaagca cgaagataca gcggtctatt actgtgctgc aagagcaccc gtcggtcaat
     4021 cttcctctcc ttatgattat gactattggg gacaggggac tcaggtaact gtgtccagtt
     4081 agacgcgtta agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     4141 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     4201 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     4261 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     4321 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     4381 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     4441 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     4501 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     4561 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     4621 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     4681 ccccgcgtcg actttaagac caatgactta caaggcagct gtagatctta gccacttttt
     4741 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt
     4801 tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     4861 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     4921 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     4981 aatctctagc agtacgtata gtagttcatg tcatcttatt attcagtatt tataacttgc
     5041 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa
     5101 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5161 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta
     5221 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     5281 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     5341 tagtgaggag gcttttttgg aggcctaggg acgtacccaa ttcgccctat agtgagtcgt
     5401 attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     5461 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     5521 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct
     5581 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     5641 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     5701 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     5761 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     5821 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     5881 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     5941 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     6001 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
     6061 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     6121 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     6181 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     6241 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     6301 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     6361 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     6421 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     6481 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     6541 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     6601 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     6661 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     6721 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     6781 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6841 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     6901 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6961 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     7021 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     7081 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     7141 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     7201 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     7261 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     7321 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     7381 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     7441 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     7501 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     7561 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     7621 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     7681 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     7741 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     7801 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     7861 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     7921 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     7981 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     8041 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     8101 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     8161 cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga
     8221 acaaaagctg gagctgcaag cttaa
//
