LOCUS       sequence-377945-        8383 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rapamycin-dependent opioid sensor for the mu opioid receptor 
            (component 1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8383)
  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 8383)
  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-MOR-FRB
FEATURES             Location/Qualifiers
     source          1..8383
                     /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             2658..2681
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    4236..4824
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4289..4309)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4707..4718)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4896..5129
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5207..5328
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5244..5263)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5298..5317
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5368..5503
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5523..5542)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5524..5542)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5552..5569)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5552..5568)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5561..5583)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5710..6165
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5797..5816)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6007..6028
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6191..6295
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6296..6364,6365..7156)
                     /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(6514..6533)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7327..7915
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7816..7835
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8069..8086
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8203..8224
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8239..8244,8245..8262,8263..8269)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8277..8293
                     /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     8282..8304
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8301..8317
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8301..8317
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8336..8356
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8338..8356
                     /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 tgaagaccat catcgccctg agctacatct
     2641 tctgcctggt gttcgccgac tacaaggatg acgacgataa gggcggaatg gactcttccg
     2701 ctgctcccac caatgcctca aattgtacag acgccttggc ctatagctct tgctctcccg
     2761 caccatctcc gggaagctgg gttaacctca gtcatctcga cgggaacctg tccgaccctt
     2821 gtggccctaa taggaccgac ctcggtggtc gagatagcct gtgcccccct actggaagcc
     2881 ccagcatgat cacagctatc acaataatgg ctctctatag tatcgtctgc gtcgtgggac
     2941 tctttggcaa ttttctcgtc atgtacgtta tcgtcagata cacaaagatg aaaaccgcaa
     3001 ccaatattta catctttaat ctggccctgg cggatgccct tgcaaccagt acactgccgt
     3061 ttcaatccgt gaattatctg atgggcacat ggcccttcgg aaccattctg tgcaaaatcg
     3121 taatctcaat tgattattat aacatgttca cctccatttt taccttgtgt acaatgtcag
     3181 tagaccgcta tatcgccgta tgccaccctg tcaaagctct tgatttcagg acccctagga
     3241 atgccaagat catcaatgtg tgcaattgga ttctgtctag cgctatcggc ttgccggtga
     3301 tgttcatggc cactaccaag tacagacaag gttcaatcga ctgtaccctc acattttcac
     3361 atcccacgtg gtattgggag aacttgctga agatctgtgt gttcatcttc gccttcatca
     3421 tgccagtact gataatcacc gtttgttatg gtcttatgat cctgcgcctg aagagcgtgc
     3481 gcatgctcag cggctccaaa gaaaaggacc ggaaccttcg gcgaatcaca cggatggtcc
     3541 tggtggttgt ggctgtgttc attgtttgct ggacccctat tcatatttat gttataatta
     3601 aggccctggt gaccatcccg gaaaccacat tccagaccgt ttcctggcac ttttgtatag
     3661 cattgggata taccaacagt tgccttaatc ccgttctgta tgcttttctg gacgagaatt
     3721 tcaagcgatg cttcagggag ttctgcattc ccacaagctc taacattgaa cagcagaatt
     3781 ctacacgaat acgacagaat accagagatc atcctagcac tgccaatact gtcgaccgga
     3841 ccaatcacca gcttgagaac cttgaggcag aaaccgcccc tctcccagag ctggccgaga
     3901 agctggccgg cctggacatc aacggtggag ctagcgtggc catcctgtgg cacgagatgt
     3961 ggcatgaagg cctggaagag gcatctcgtt tgtactttgg ggaaaggaac gtgaaaggca
     4021 tgtttgaggt gctggagccc ttgcatgcta tgatggaacg gggcccccag actctgaagg
     4081 aaacatcctt taatcaggcc tatggtcgag atttaatgga ggcccaagag tggtgcagga
     4141 agtacatgaa atcagggaat gtcaaggacc tcacccaagc ctgggacctc tattatcatg
     4201 tgttccgacg aatctcatag acgcgttaag tcgacaatca acctctggat tacaaaattt
     4261 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     4321 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     4381 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     4441 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     4501 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     4561 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     4621 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     4681 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     4741 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     4801 tctccctttg ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt
     4861 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     4921 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
     4981 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     5041 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     5101 cttttagtca gtgtggaaaa tctctagcag tacgtatagt agttcatgtc atcttattat
     5161 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc
     5221 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5281 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct
     5341 ctagctatcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     5401 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     5461 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt
     5521 cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact
     5581 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     5641 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     5701 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     5761 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     5821 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     5881 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     5941 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     6001 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     6061 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     6121 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     6181 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     6241 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     6301 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     6361 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     6421 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     6481 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     6541 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     6601 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     6661 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     6721 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     6781 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     6841 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     6901 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6961 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     7021 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     7081 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     7141 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     7201 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     7261 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     7321 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     7381 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     7441 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     7501 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     7561 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     7621 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     7681 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     7741 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     7801 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     7861 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     7921 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     7981 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     8041 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     8101 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     8161 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     8221 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     8281 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat
     8341 taaccctcac taaagggaac aaaagctgga gctgcaagct taa
//
