LOCUS       sequence-467730-        7954 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRET-based intramolecular conformation sensor for human KOR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7954)
  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  JOURNAL   Unpublished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 7954)
  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            "Unpublished"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: KOR FRET sensor
FEATURES             Location/Qualifiers
     source          1..7954
                     /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             complement(1594..1605)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             join(1750..1752,1753..1755,1756..2466)
                     /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(2785..2787,2788..3498)
                     /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(2785..2787,2788..3498)
                     /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(3545..3562)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3551..3775
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3821..4249
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3908..3927)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4118..4139
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4258..4278)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4263..4592
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4443..4578
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4505..4524
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4659..5453
                     /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(4713..4732)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5323..5342
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5627..5748
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5664..5683)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5718..5737
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5797..5813)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5797..5813)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5810..5832)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5821..5837
                     /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(5845..5851,5852..5869,5870..5875))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5890..5911
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6028..6045)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6199..6787)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6279..6298)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6958..7749,7750..7818))
                     /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     7581..7600
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7819..7923)
                     /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 atggaatccc cgattcagat cttccgcggg gagccgggcc ctacctgcgc cccgagcgcc
     1021 tgcctgcccc ccaacagcag cgcctggttt cccggctggg ccgagcccga cagcaacggc
     1081 agcgccggct cggaggacgc gcagctggag cccgcgcaca tctccccggc catcccggtc
     1141 atcatcacgg cggtctactc cgtagtgttc gtcgtgggct tggtgggcaa ctcgctggtc
     1201 atgttcgtga tcatccgata cacaaagatg aagacagcaa ccaacattta catatttaac
     1261 ctggctttgg cagatgcttt agttactaca accatgccct ttcagagtac ggtctacttg
     1321 atgaattcct ggccttttgg ggatgtgctg tgcaagatag taatttccat tgattactac
     1381 aacatgttca ccagcatctt caccttgacc atgatgagcg tggaccgcta cattgccgtg
     1441 tgccaccccg tgaaggcttt ggacttccgc acacccttga aggcaaagat catcaatatc
     1501 tgcatctggc tgctgtcgtc atctgttggc atctctgcaa tagtccttgg aggcaccaaa
     1561 gtcagggaag acgtcgatgt cattgagtgc tccttgcagt tcccagatga tgactactcc
     1621 tggtgggacc tcttcatgaa gatctgcgtc ttcatctttg ccttcgtgat ccctgtcctc
     1681 atcatcatcg tctgctacac cctgatgatc ctgcgtctca agagcgtccg gctcctttct
     1741 ggcccgcgga tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     1801 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     1861 gccacctacg gcaagctgac cctgaagctg atctgcacca ccggcaagct gcccgtgccc
     1921 tggcccaccc tcgtgaccac cctgggctac ggcgtgcagt gcttcgcccg ctaccccgac
     1981 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     2041 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     2101 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     2161 ctggggcaca agctggagta caactacaac agccacaacg tctatatcac cgccgacaag
     2221 cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg
     2281 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     2341 gacaaccact acctgagcta ccagtccaag ctgagcaaag accccaacga gaagcgcgat
     2401 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     2461 tacaaggtta actcccgaga gaaagatcgc aacctgcgta ggatcaccag actggtcctg
     2521 gtggtggtgg cagtcttcgt cgtctgctgg actcccattc acatattcat cctggtggag
     2581 gctctgggga gcacctccca cagcacagct gctctctcca gctattactt ctgcatcgcc
     2641 ttaggctata ccaacagtag cctgaatccc attctctacg cctttcttga tgaaaacttc
     2701 aagcggtgtt tccgggactt ctgctttcca ctgaagatga ggatggagcg gcagagcact
     2761 agcagagtcc gaaatacaac cggtgtgagc aagggcgagg agctgttcac cggggtggtg
     2821 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     2881 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     2941 ctgcccgtgc cctggcccac cctcgtgacc accctgtcct ggggcgtgca gtgcttcgcc
     3001 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     3061 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     3121 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     3181 gacggcaaca tcctggggca caagctggag tacaactact tcagcgacaa cgtctatatc
     3241 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
     3301 gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     3361 gtgctgctgc ccgacaacca ctacctgagc acccagtcca agctgagcaa agaccccaac
     3421 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     3481 atggacgagc tgtacaagga tatctagctc gagtctagag ggcccgttta aacccgctga
     3541 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     3601 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     3661 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     3721 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     3781 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     3841 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3901 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3961 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4021 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4081 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4141 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     4201 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     4261 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4321 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4381 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4441 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4501 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     4561 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     4621 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4681 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4741 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4801 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4861 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4921 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4981 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5041 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5101 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5161 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     5221 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5281 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5341 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5401 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5461 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5521 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5581 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     5641 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5701 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5761 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5821 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5881 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5941 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6001 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6061 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6121 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6181 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6241 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6301 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6361 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6421 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6481 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6541 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6601 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6661 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6721 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6781 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6841 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6901 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6961 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7021 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7081 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7141 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7201 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7261 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7321 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7381 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7441 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7501 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7561 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7621 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7681 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7741 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7801 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7861 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7921 gcgcacattt ccccgaaaag tgccacctga cgtc
//
