LOCUS       sequence-467728-        7975 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRET-based intramolecular conformation sensor for rat MOR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7975)
  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  JOURNAL   Unpublished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 7975)
  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            "Unpublished"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: MOR FRET sensor
FEATURES             Location/Qualifiers
     source          1..7975
                     /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(1768..1770,1771..1773,1774..2484)
                     /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(2806..2808,2809..3519)
                     /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(2806..2808,2809..3519)
                     /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(3566..3583)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3572..3796
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3842..4270
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3929..3948)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4139..4160
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4279..4299)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4284..4613
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4464..4599
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4526..4545
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4680..5474
                     /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(4734..4753)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5344..5363
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5648..5769
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5685..5704)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5739..5758
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5818..5834)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5818..5834)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5831..5853)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5842..5858
                     /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(5866..5872,5873..5890,5891..5896))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5911..5932
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6049..6066)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6220..6808)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6300..6319)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6979..7770,7771..7839))
                     /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     7602..7621
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7840..7944)
                     /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 atggacagca gcaccggccc agggaacacc agcgactgct cagacccctt agctcaggca
     1021 agttgctccc cagcacctgg ctcctggctc aacttgtccc acgttgatgg caaccagtcc
     1081 gatccatgcg gtctgaaccg caccgggctt ggcgggaacg acagcctgtg ccctcagacc
     1141 ggcagccctt ccatggtcac agccattacc atcatggccc tctactctat cgtgtgtgta
     1201 gtgggcctct tcggaaactt cctggtcatg tatgtgattg taagatacac caaaatgaag
     1261 actgccacca acatctacat tttcaacctt gctctggcag acgccttagc gaccagtaca
     1321 ctgccctttc agagtgtcaa ctacctgatg ggaacatggc ccttcggaac catcctctgc
     1381 aagatcgtga tctcaataga ttactacaac atgttcacca gcatattcac cctctgcacc
     1441 atgagcgtgg accgctacat tgctgtctgc cacccagtca aagccctgga tttccgtacc
     1501 ccccgaaatg ccaaaatcgt caacgtctgc aactggatcc tctcttctgc catcggtctg
     1561 cctgtaatgt tcatggcaac cacaaaatac aggcaggggt ccatagattg caccctcacg
     1621 ttctcccacc caacctggta ctgggagaac ctgctcaaaa tctgtgtctt tatcttcgct
     1681 ttcatcatgc cgatcctcat catcactgtg tgttacggcc tgatgatctt acgactcaag
     1741 agcgttcgca tgctatcggg cccgcggatg gtgagcaagg gcgaggagct gttcaccggg
     1801 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     1861 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc
     1921 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg cgtgcagtgc
     1981 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     2041 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     2101 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     2161 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     2221 tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac
     2281 atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     2341 ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccaagct gagcaaagac
     2401 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     2461 ctcggcatgg acgagctgta caaggttaac tccaaagaaa aggacaggaa tctgcgcagg
     2521 atcacccgga tggtgctggt ggtcgtggct gtatttatcg tctgctggac ccccatccac
     2581 atctacgtca tcatcaaagc gctgatcacg attccagaaa ccacatttca gaccgtttcc
     2641 tggcacttct gcattgcttt gggttacacg aacagctgcc tgaatccagt tctttacgcc
     2701 ttcctggatg aaaacttcaa gcgatgcttc agagagttct gcatcccaac ctcgtccacg
     2761 atcgaacagc aaaactccac tcgagtccgt cagaacacta ccggtgtgag caagggcgag
     2821 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     2881 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     2941 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgtcc
     3001 tggggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     3061 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     3121 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     3181 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     3241 ttcagcgaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc
     3301 aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac
     3361 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     3421 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     3481 gccgccggga tcactctcgg catggacgag ctgtacaagg atatctagct cgagtctaga
     3541 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3601 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3661 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3721 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3781 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     3841 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3901 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3961 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4021 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     4081 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4141 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4201 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4261 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4321 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4381 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4441 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4501 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     4561 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     4621 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     4681 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4741 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4801 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4861 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4921 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4981 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     5041 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     5101 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     5161 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     5221 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     5281 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5341 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5401 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5461 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     5521 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5581 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5641 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5701 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5761 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5821 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5881 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5941 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6001 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6061 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6121 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6181 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6241 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6301 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6361 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     6421 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6481 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6541 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6601 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6661 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6721 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6781 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6841 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6901 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6961 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7021 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7081 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7141 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7201 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7261 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7321 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7381 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7441 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7501 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7561 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7621 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7681 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7741 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7801 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7861 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7921 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
