LOCUS       sequence-469825-        7675 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human B2-adrenergic receptor fused to RLuc8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7675)
  AUTHORS   Gimenez LE, Vishnivetskiy SA, Baameur F, Gurevich VV
  TITLE     Manipulation of very few receptor discriminator residues greatly 
            enhances receptor specificity of non-visual arrestins.
  JOURNAL   J Biol Chem. 2012 Aug 24;287(35):29495-505. doi: 
            10.1074/jbc.M112.366674. Epub 2012 Jul 11.
  PUBMED    22787152
REFERENCE   2  (bases 1 to 7675)
  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: 
            "10.1074/jbc.M112.366674"; journalName: "J Biol Chem"; date: 
            "2012-08-24- 24"; volume: "287"; issue: "35"; pages: "29495-505"
COMMENT     Sequence Label: p3HB2ALuc
FEATURES             Location/Qualifiers
     source          1..7675
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(148..167)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      961..1549
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1450..1469
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1703..1720
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1837..1858
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1873..1878,1879..1896,1897..1903)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1911..1927
                     /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     1916..1938
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1935..1951
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1935..1951
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2000..2121
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2011..2030)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2065..2084
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2295..3089)
                     /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(2406..2425)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3016..3035
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3156..3485)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3170..3305
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3224..3243)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     3470..3490
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(3499..3927)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3609..3630)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3821..3840
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3973..4197
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4186..4203
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        4223..4241
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4223..4240
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(4280..5215)
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNATSSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGAALAFHYAYEHQDRIKAIVHMESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETVLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKLFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFLQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             complement(6467..6547)
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     primer_bind     complement(6587..6606)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6588..6606)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(6651..6854)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6680..6700)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6855..7234
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     7406..7425
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        7500..7604
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7605..7673,7674..7675,1..790)
                     /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"
ORIGIN
        1 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
       61 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
      121 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
      181 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
      241 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
      301 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
      361 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
      421 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
      481 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
      541 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
      601 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
      661 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
      721 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
      781 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
      841 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
      901 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
      961 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1021 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     1081 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     1141 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     1201 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     1261 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     1321 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     1381 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     1441 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     1501 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     1561 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     1621 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     1681 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     1741 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     1801 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     1861 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     1921 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt
     1981 cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     2041 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     2101 ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat ccccgcgctg
     2161 gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc
     2221 ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg tcatttcgaa
     2281 ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa
     2341 tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct
     2401 tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac acccagccgg
     2461 ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca
     2521 tcgccatggg tcacgacgag atcctcgccg tcgggcatgc gcgccttgag cctggcgaac
     2581 agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg
     2641 gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag
     2701 gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg
     2761 gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa tagcagccag
     2821 tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc
     2881 agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga caggtcggtc
     2941 ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag
     3001 ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa
     3061 cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatca
     3121 gatccgaaaa tggatataca agctcccggg agctttttgc aaaagcctag gcctccaaaa
     3181 aagcctcctc actacttctg gaatagctca gaggcagagg cggcctcggc ctctgcataa
     3241 ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc
     3301 gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc
     3361 atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga
     3421 tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga
     3481 cacacattcc acagaattaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     3541 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     3601 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     3661 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     3721 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     3781 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     3841 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     3901 cgcttaatgc gccgctacag ggcgcgtggg gataccccct agagccccag ctggttcttt
     3961 ccgcctcaga agccatagag cccaccgcat ccccagcatg cctgctattg tcttcccaat
     4021 cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac ctactcagac
     4081 aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca ccttccaggg
     4141 tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa ggcacagtcg
     4201 aggctgatca gcgagctcta gcatttaggt gacactatag aatagggcca agcttttaat
     4261 taatctagag gcgcgccgat tactgctcgt tcttcagcac gcgctccacg aagctcttga
     4321 tgtacttacc catttcatct ggagcgtcct cctggaggaa gtggaggccc ttcaccttca
     4381 cgaactcggt gttagggaac ttcttagctc cctcgacaat agcgttggaa aagaacccag
     4441 ggtcggactc gatgaacagc ttaggcagat cgtcgctggc ccgaaggtag gcgttgtagt
     4501 tgcggacaat ctggacgacg tcgggcttgc ctcccttaac gagagggatc tcgcgaggcc
     4561 aggagagggt aggccgtcta acctcgccct tctccttgaa tggctccagg taggcagcga
     4621 actcctcagg ctccagtttc cgcatgatct tgcttgggag cacggtctcg acgaagaagt
     4681 tattctcaag caccattttc tcgccctctt cgctcttgat cagggcgata tcctcctcga
     4741 tgtcaggcca ctcgtcccag gactcgatca cgtccacgac actctccata tggacgatgg
     4801 ccttgatcct gtcttggtgc tcgtaggcgt agtgaaaggc cagagcagcc ccccagtcgt
     4861 ggcccacaaa gatgattttc tttggaaggt tcagcagctc gaaccaagcg gtgaggtact
     4921 tgtagtgatc caggaggcga tatgagccat tcccgctctt gccggactta cccattccga
     4981 tcagatcagg gatgatgcat ctagccacgg gctcgatgtg aggcacgacg tgcctccaca
     5041 ggtagctgga ggtagcgtta ccatgcagaa aaatcacggc gttctcggcg tgcttctcgg
     5101 aatcatagta gttgatgaag gagtccagca cgttcatttg cttgcagcga gcccaccact
     5161 gaggcccagt gatcatgcgt ttgcgttgct cggggtcgta caccttggaa gccatgcctg
     5221 caggtgccag cagtgagtca tttgtactac aattcctccc ttgtgaatca atgttatcgc
     5281 taggcacagt accttgatgg cccacaaagt cttccgtgcc tgggaggtct tcacacagca
     5341 gtttattttc tttctcctgt tccacgtgat atccactctg ctcccctgtg ttgccgttgc
     5401 tggagtagcc attcccatag gccttcaaag aagacctgcg caggcacaga agctcctgga
     5461 aggcaatcct gaaatctggg ctccggcagt agataagggg attgaaacca gaattgacat
     5521 agcctatcca atttaggagg atgtaaactt ccttacggat gaggttatcc tggatcacat
     5581 gcacaatgtt aacgatgaag aagggcagcc agcagagggt gaaagtgccc atgatgatgc
     5641 ctaacgtctt gagggctttg tgctccttca agcagaactt ggaagatctg cggagtccat
     5701 gccccgtccg cccatcctgc tccacctggc taaggttctg gacatggaag cggccctcag
     5761 atttgtcaat cttctggagc tgccttttgg cctcctgaaa gaccctggag tagacgaaga
     5821 ccatgatcac caggggaacg tagaaggaca cgatggaaga ggcaatggca taggcttggt
     5881 tcgtgaagaa gtcacagcag gtctcattgg catagcagtt gatggcttcc tggtgggtgg
     5941 cccggtacca gtgcatctga atgggcaaga aggaggtaag gcctgacaca atccacacca
     6001 tcagaatgat cacccgggcc ttattcttgg tcagcaggct ctggtacttg aaaggtgaag
     6061 taatggcaaa gtagcgatcc actgcgatca cgcacagggt ctcaatgctg gccgtgacgc
     6121 acagcacatc aatggaagtc caaaactcgc accagaagtt gccaaaagtc cacattttca
     6181 taagaatatg ggcggcccca aagggcacca ctgccaggcc catgaccaga tcagcacagg
     6241 ccagtgaagt gatgaagtag ttggtgaccg tctgcagacg ctcgaacttg gcaatggctg
     6301 tgatgaccag cacattgcca aacacgatgg ccaggacgat gagagacatg acgatgccca
     6361 tgcccaccac ccacacctcg tccctttgct gcgtgacgtc gtggtccggc gcatggcttc
     6421 tattgggtgc cagcaagaag gcgctgccgt tcccgggttg cccatcagcg taatctggaa
     6481 catcgtatgg gtaagcgtaa tctggaacat cgtatgggta agcgtaatct ggaacatcgt
     6541 atgggtacgc catggtggtg cgatcgcaga attccagctt gggtctccct atagtgagtc
     6601 gtattaattt cgataagcca gtaagcagtg ggttctctag ttagccagag agctctgctt
     6661 atatagacct cccaccgtac acgcctaccg cccatttgcg tcaatggggc ggagttgtta
     6721 cgacattttg gaaagtcccg ttgattttgg tgccaaaaca aactcccatt gacgtcaatg
     6781 gggtggagac ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt gatgtactgc
     6841 caaaaccgca tcaccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga
     6901 aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg
     6961 tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac
     7021 cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca
     7081 tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt
     7141 accgtaagtt atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt
     7201 gattactatt aataactagt caataatcaa tgtcaacgcg tatatctggc ccgtacatcg
     7261 cgaagcagcg caaaacgcct aaccctaagc agattcttca tgcaattgtc ggtcaagcct
     7321 tgccttgttg tagcttaaat tttgctcgcg cactactcag cgacctccaa cacacaagca
     7381 gggagcagat actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     7441 aggggatcgg gagatctccc gatccgtcga cgtcaggtgg cacttttcgg ggaaatgtgc
     7501 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     7561 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     7621 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctca
//
