LOCUS       sequence-377855-        8317 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rapamycin-dependent opioid sensor for the kappa opioid receptor 
            (component 1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8317)
  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 8317)
  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-KOR-FRB
FEATURES             Location/Qualifiers
     source          1..8317
                     /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"
     misc_feature    4174..4762
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4227..4247)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4645..4656)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4834..5067
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5145..5266
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5182..5201)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5236..5255
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5306..5441
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5461..5480)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5462..5480)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5490..5507)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5490..5506)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5499..5521)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5648..6103
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5735..5754)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5945..5966
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6129..6233
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6234..6302,6303..7094)
                     /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(6452..6471)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7265..7853
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7754..7773
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8007..8024
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8141..8162
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8177..8182,8183..8200,8201..8207)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8215..8231
                     /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     8220..8242
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8239..8255
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8239..8255
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8274..8294
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8276..8294
                     /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 accgctagca tgaagacgat catcgccctg
     2641 agctacatct tctgcctggt gttcgccgac tacaaggacg atgatgacgc catggacagc
     2701 cccattcaaa tcttccgggg cgagcccgga ccaacctgcg ctccaagcgc ttgcctgcca
     2761 ccgaattcta gtgcatggtt cccagggtgg gcagagcccg atagcaatgg ctcagccggg
     2821 tccgaggatg cgcagcttga acccgctcac atctcacccg ctatccctgt cattattacg
     2881 gcagtctact cagttgtttt cgttgtgggc ctggttggaa atagcctcgt gatgtttgtg
     2941 attatcaggt atactaaaat gaagaccgcc acgaatatct acatctttaa cctggctctg
     3001 gctgacgcct tggtgaccac cactatgcca tttcagtcca ccgtctatct tatgaactct
     3061 tggccattcg gcgacgttct gtgcaagatt gtcatttcca ttgactatta taatatgttt
     3121 acatcaatct ttacgcttac aatgatgagc gtggaccgat acattgctgt ttgccatccg
     3181 gtgaaagcac tggattttcg cacccccttg aaggccaaga ttatcaatat ctgtatttgg
     3241 ctcctgtcct catccgtcgg aatttccgcc attgtcttgg gtggtacaaa ggtgagggag
     3301 gacgtggacg ttatagagtg cagcctgcag ttccctgacg atgactacag ctggtgggac
     3361 ctttttatga agatctgtgt cttcattttt gctttcgtca tccccgtgct gattatcata
     3421 gtgtgctata cactgatgat attgcggctg aaatctgtgc gcctgctgag cggcagccgc
     3481 gaaaaggatc ggaatctgcg ccgaatcaca cgcctcgtgc tggttgtagt cgctgtcttc
     3541 gtggtatgct ggaccccaat ccacattttc atcctggtgg aagctctggg atccacctct
     3601 cactccactg ccgcacttag ctcatattac ttctgcattg ccttgggcta cactaattcc
     3661 tcactgaatc ccattctgta cgcatttctt gacgagaact tcaaacggtg ctttagagac
     3721 ttttgcttcc ccctcaagat gcgcatggag cggcagtcaa ccagccgagt ccggaatacg
     3781 gtccaggatc ccgcctatct gcgggacatc gacggcatga acaagcctgt gcagggcgcg
     3841 cccaagggct cgggctcgac ctcgggctcg ggcgtggcca tcctgtggca cgagatgtgg
     3901 catgaaggcc tggaagaggc atctcgtttg tactttgggg aaaggaacgt gaaaggcatg
     3961 tttgaggtgc tggagccctt gcatgctatg atggaacggg gcccccagac tctgaaggaa
     4021 acatccttta atcaggccta tggtcgagat ttaatggagg cccaagagtg gtgcaggaag
     4081 tacatgaaat cagggaatgt caaggacctc acccaagcct gggacctcta ttatcatgtg
     4141 ttccgacgaa tctcatagac gcgttaagtc gacaatcaac ctctggatta caaaatttgt
     4201 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgatgct
     4261 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     4321 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     4381 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     4441 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     4501 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     4561 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     4621 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     4681 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     4741 tccctttggg ccgcctcccc gcgtcgactt taagaccaat gacttacaag gcagctgtag
     4801 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     4861 gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     4921 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     4981 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     5041 tttagtcagt gtggaaaatc tctagcagta cgtatagtag ttcatgtcat cttattattc
     5101 agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag
     5161 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5221 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct
     5281 agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     5341 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     5401 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt acccaattcg
     5461 ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg
     5521 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     5581 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     5641 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     5701 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     5761 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     5821 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     5881 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     5941 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     6001 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     6061 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     6121 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     6181 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     6241 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     6301 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     6361 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     6421 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     6481 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     6541 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     6601 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     6661 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     6721 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     6781 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     6841 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     6901 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     6961 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     7021 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     7081 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     7141 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     7201 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     7261 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     7321 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     7381 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     7441 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     7501 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     7561 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     7621 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     7681 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     7741 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     7801 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     7861 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     7921 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     7981 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     8041 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     8101 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     8161 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     8221 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta
     8281 accctcacta aagggaacaa aagctggagc tgcaagc
//
