LOCUS       sequence-467739-        7341 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminally sYFP-tagged Arrestin3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7341)
  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  JOURNAL   Unpublished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 7341)
  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            "Unpublished"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: sYFP-Arrestin3
FEATURES             Location/Qualifiers
     source          1..7341
                     /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(895..897,898..900,901..1611)
                     /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"
     primer_bind     complement(940..961)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1201..1220)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1548..1569
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(2894..2912)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2895..2912)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2932..2949)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2938..3162
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3208..3636
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3295..3314)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3505..3526
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3645..3665)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3650..3979
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3830..3965
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3892..3911
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4046..4840
                     /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(4100..4119)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4710..4729
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5014..5135
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5051..5070)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5105..5124
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5184..5200)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5184..5200)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5197..5219)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5208..5224
                     /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(5232..5238,5239..5256,5257..5262))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5277..5298
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5415..5432)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5586..6174)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5666..5685)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6345..7136,7137..7205))
                     /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     6968..6987
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7206..7310)
                     /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 gcttatggtg
      901 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
      961 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     1021 ctgaccctga agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     1081 accaccctgg gctacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
     1141 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     1201 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1261 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1321 gagtacaact acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc
     1381 aaggccaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac
     1441 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1501 agctaccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1561 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa ggaattctgc
     1621 agatatcaga tgggggagaa acccgggacc agggtcttca agaaatcgag tcctaactgc
     1681 aagctcaccg tgtacctggg caagcgggac tttgtggacc acctggacaa agtggatcct
     1741 gtagatggtg tggtgctagt ggacccggac tacttgaagg accgcaaagt gtttgtgacc
     1801 ctcacctgcg cctttcgcta tggccgcgaa gacctggacg tgctgggctt gtccttccgc
     1861 aaagacctgt tcatcgccaa ctaccaggcc ttccccccaa cacccaaccc accccggccc
     1921 cccacccgcc tgcaggagcg gctgctgagg aagctgggcc agcatgccca cccctttttt
     1981 ttcacaatac cccagaacct gccctgctcc gtcaccctgc agccaggccc agaggatacg
     2041 gggaaggcct gcggggtaga ctttgagatt cgagccttct gcgccaaatc actagaagag
     2101 aaaagccaca aacggaactc tgtgcgtctg gtgatccgaa aggtgcagtt tgccccagag
     2161 aaacccggcc cccagccttc agctgaaacc acacgccact tcctcatgtc tgaccggtcc
     2221 ctgcaccttg aggcttcact ggacaaggag ctgtactacc atggggagcc cctcaatgtc
     2281 aatgtccacg tcaccaataa ctccaccaag actgtcaaga agatcaaagt ctctgtgaga
     2341 cagtacgccg acatctgcct cttcagcacc gcccagtaca agtgccccgt ggctcaggtc
     2401 gaacaagatg accaggtgtc gcccagttcc acgttctgta aggtgtacac cataaccccg
     2461 ctgctcagca acaaccggga gaagcgcggc ctcgctctgg atgggaagct caagcacgag
     2521 gacaccaacc tggcgtccag caccattgtg aaggagggtg ccaacaagga ggtgctgggg
     2581 atcctggtgt cctacagggt caaggtgaag ctggtggtgt ctcgaggcgg ggacgtctca
     2641 gtggagctgc catttgttct catgcacccc aagccccatg atcacatcgc ccttcccagg
     2701 ccccagtcag ctgttcctga aacagatgcc cccgtggaca ccaacctcat tgaattcgaa
     2761 accaactatg ccacagacga cgacatcgtg tttgaggact ttgcccggct gcggctcaag
     2821 gggctgaagg acgaggacta tgacgaccag ttctgttaag cggccgctcg agcatgcatc
     2881 tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg
     2941 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     3001 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     3061 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     3121 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     3181 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     3241 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3301 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3361 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3421 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3481 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     3541 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     3601 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     3661 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3721 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3781 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3841 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3901 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     3961 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     4021 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4081 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4141 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4201 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     4261 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4321 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4381 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4441 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4501 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4561 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4621 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4681 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4741 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4801 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     4861 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4921 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4981 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     5041 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     5101 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     5161 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     5221 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     5281 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5341 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5401 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5461 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5521 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5581 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5641 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5701 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5761 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5821 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5881 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5941 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6001 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     6061 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6121 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6181 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6241 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6301 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6361 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6421 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6481 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     6541 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6601 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6661 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6721 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6781 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6841 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6901 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6961 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     7021 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     7081 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7141 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7201 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7261 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7321 cgaaaagtgc cacctgacgt c
//
