LOCUS       sequence-467736-        7918 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRET-based intramolecular conformation sensor for human nociceptin 
            receptor NOP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7918)
  TITLE     Dynamics of agonist-evoked opioid receptor activation revealed by 
            FRET- and BRET-based opioid receptor conformation sensors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7918)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NOP FRET sensor
FEATURES             Location/Qualifiers
     source          1..7918
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(1717..1719,1720..1722,1723..2433)
                     /codon_start=1
                     /product="monomeric optimized variant of EYFP, identical to
                     mVenus but with L68V (Kremers et al., 2006)"
                     /label=SYFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2749..2751,2752..3462)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2749..2751,2752..3462)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2010)"
                     /label=mTurquoise
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3509..3526)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3515..3739
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3785..4213
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3872..3891)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4082..4103
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4222..4242)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4227..4556
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4407..4542
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4469..4488
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4623..5417
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4677..4696)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5287..5306
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5591..5712
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5628..5647)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5682..5701
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5761..5777)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5761..5777)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5774..5796)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5785..5801
                     /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)."
     promoter        complement(join(5809..5815,5816..5833,5834..5839))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5854..5875
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5992..6009)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6163..6751)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6243..6262)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6922..7713,7714..7782))
                     /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     7545..7564
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7783..7887)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcacc
      961 atggagcccc tgtttccggc ccccttttgg gaagtcatct atggctcaca tctgcagggg
     1021 aacctgagtc tgctgtcccc caaccattca ctgcttcctc cccatctgtt gctgaacgct
     1081 tctcatggag cattcctgcc tctcggattg aaggtaacta tcgtcggctt gtatctcgcc
     1141 gtctgcgtgg gcggcctcct gggtaactgt ctggttatgt acgttattct ccggcacact
     1201 aaaatgaaga cagcaacaaa catttacatc tttaatctgg cactggctga tacactggta
     1261 ttgttgacgc tgcctttcca ggggaccgat attctcctgg gattctggcc tttcgggaac
     1321 gccctgtgta agactgtaat agcaatagat tattacaaca tgtttactag cacattcacg
     1381 ctgacagcca tgagtgtgga tagatacgtg gctatctgcc accccattcg agcactggac
     1441 gtgcgcacct cctcaaaagc ccaagcagtc aatgtggcca tctgggcact ggccagtgtg
     1501 gttggtgtgc ccgtagcgat aatgggcagc gcccaagtgg aggacgaaga gatcgaatgc
     1561 ctcgtggaga ttccaactcc acaggactat tggggccctg tctttgccat ctgtatcttt
     1621 ttgttctctt tcatcgtgcc agtactcgtc atctcagtat gttacagcct gatgattcgc
     1681 aggctccgag gcgttagact gctgagtgga ccgcggatgg tgagcaaggg cgaggagctg
     1741 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     1801 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagctgatc
     1861 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gggctacggc
     1921 gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     1981 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     2041 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     2101 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     2161 cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc
     2221 cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc
     2281 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca gtccaagctg
     2341 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     2401 gggatcactc tcggcatgga cgagctgtac aaggttaaca gcagggagaa ggaccgcaac
     2461 ctccgacgca tcacgaggct ggtcctcgtg gttgtggcgg tgttcgtggg ctgttggacc
     2521 cccgtgcagg tcttcgttct cgcccaggga cttggggtgc agccgtcaag cgagacagca
     2581 gtggctatac tgcgcttttg caccgcactg gggtacgtca acagctgcct gaacccaatt
     2641 ctttacgctt ttctcgatga gaatttcaag gcctgcttcc ggaaattctg ttgtgcttca
     2701 gccctgcgac gggatgtaca agtctcagac agagtgaggt caaccggtgt gagcaagggc
     2761 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     2821 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     2881 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     2941 tcctggggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     3001 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     3061 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     3121 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     3181 tacttcagcg acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac
     3241 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag
     3301 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     3361 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     3421 accgccgccg ggatcactct cggcatggac gagctgtaca aggatatcta gctcgagtct
     3481 agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     3541 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3601 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3661 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     3721 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     3781 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3841 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3901 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3961 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4021 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4081 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4141 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4201 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4261 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4321 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4381 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4441 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     4501 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     4561 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     4621 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4681 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4741 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4801 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4861 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4921 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4981 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5041 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5101 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     5161 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5221 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5281 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5341 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5401 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     5461 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5521 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5581 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     5641 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5701 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     5761 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5821 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5881 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5941 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6001 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6061 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6121 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6181 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6241 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6301 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6361 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6421 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6481 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6541 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6601 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6661 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6721 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6781 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6841 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6901 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6961 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7021 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7081 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     7141 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     7201 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     7261 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     7321 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7381 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7441 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7501 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7561 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7621 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7681 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7741 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7801 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7861 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
