LOCUS       sequence-471673-        7774 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human dopamine D1 receptor; N-terminal HA 
            tag, C-terminal Rluc8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7774)
  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 7774)
  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: p3HD1RLuc2
FEATURES             Location/Qualifiers
     source          1..7774
                     /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             922..1002
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             2353..3288
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNATSSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGAALAFHYAYEHQDRIKAIVHMESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETVLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKLFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFLQEDAPDEMGKYIKSFVERVLKNEQ"
     promoter        complement(3327..3345)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3328..3345)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3365..3382)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3371..3595
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3641..4069
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3728..3747)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3938..3959
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4078..4098)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4083..4412
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4263..4398
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4325..4344
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4479..5273
                     /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(4533..4552)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5143..5162
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5447..5568
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5484..5503)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5538..5557
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5617..5633)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5617..5633)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5630..5652)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5641..5657
                     /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(5665..5671,5672..5689,5690..5695))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5710..5731
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5848..5865)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6019..6607)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6099..6118)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6778..7569,7570..7638))
                     /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     7401..7420
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7639..7743)
                     /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 gctggaattc
      901 tgcgatcgca ccaccatggc gtacccatac gatgttccag attacgctta cccatacgat
      961 gttccagatt acgcttaccc atacgatgtt ccagattacg ctgataggac tctgaacacc
     1021 tctgccatgg acgggactgg gctggtggtg gagagggact tctctgttcg tatcctcact
     1081 gcctgtttcc tgtcgctgct catcctgtcc acgctcctgg ggaacacgct ggtctgtgct
     1141 gccgttatca ggttccgaca cctgcggtcc aaggtgacca acttctttgt catctccttg
     1201 gctgtgtcag atctcttggt ggccgtcctg gtcatgccct ggaaggcagt ggctgagatt
     1261 gctggcttct ggccctttgg gtccttctgt aacatctggg tggcctttga catcatgtgc
     1321 tccactgcat ccatcctcaa cctctgtgtg atcagcgtgg acaggtattg ggctatctcc
     1381 agccctttcc ggtatgagag aaagatgacc cccaaggcag ccttcatcct gatcagtgtg
     1441 gcatggacct tgtctgtact catctccttc atcccagtgc agctcagctg gcacaaggca
     1501 aaacccacaa gcccctctga tggaaatgcc acttccctgg ctgagaccat agacaactgt
     1561 gactccagcc tcagcaggac atatgccatc tcatcctctg taataagctt ttacatccct
     1621 gtggccatca tgattgtcac ctacaccagg atctacagga ttgctcagaa acaaatacgg
     1681 cgcattgcgg ccttggagag ggcagcagtc cacgccaaga attgccagac caccacaggt
     1741 aatggaaagc ctgtcgaatg ttctcaaccg gaaagttctt ttaagatgtc cttcaaaaga
     1801 gaaactaaag tcctgaagac tctgtcggtg atcatgggtg tgtttgtgtg ctgttggcta
     1861 cctttcttca tcttgaactg cattttgccc ttctgtgggt ctggggagac gcagcccttc
     1921 tgcattgatt ccaacacctt tgacgtgttt gtgtggtttg ggtgggctaa ttcatccttg
     1981 aaccccatca tttatgcctt taatgctgat tttcggaagg cattttcaac cctcttagga
     2041 tgctacagac tttgccctgc gacgaataat gccatagaga cggtgagtat caataacaat
     2101 ggggccgcga tgttttccag ccatcatgag ccacgaggct ccatctccaa ggagtgcaat
     2161 ctggtttacc tgatcccaca tgctgtgggc tcctctgagg acctgaaaaa ggaggaggca
     2221 gctggcatcg ccagaccctt ggagaagctg tccccagccc tatcggtcat attggactat
     2281 gacactgacg tctctctgga gaagatccaa cccatcacac aaaacggtca gcacccaacc
     2341 gcacctgcag gcatggcttc caaggtgtac gaccccgagc aacgcaaacg catgatcact
     2401 gggcctcagt ggtgggctcg ctgcaagcaa atgaacgtgc tggactcctt catcaactac
     2461 tatgattccg agaagcacgc cgagaacgcc gtgatttttc tgcatggtaa cgctacctcc
     2521 agctacctgt ggaggcacgt cgtgcctcac atcgagcccg tggctagatg catcatccct
     2581 gatctgatcg gaatgggtaa gtccggcaag agcgggaatg gctcatatcg cctcctggat
     2641 cactacaagt acctcaccgc ttggttcgag ctgctgaacc ttccaaagaa aatcatcttt
     2701 gtgggccacg actggggggc tgctctggcc tttcactacg cctacgagca ccaagacagg
     2761 atcaaggcca tcgtccatat ggagagtgtc gtggacgtga tcgagtcctg ggacgagtgg
     2821 cctgacatcg aggaggatat cgccctgatc aagagcgaag agggcgagaa aatggtgctt
     2881 gagaataact tcttcgtcga gaccgtgctc ccaagcaaga tcatgcggaa actggagcct
     2941 gaggagttcg ctgcctacct ggagccattc aaggagaagg gcgaggttag acggcctacc
     3001 ctctcctggc ctcgcgagat ccctctcgtt aagggaggca agcccgacgt cgtccagatt
     3061 gtccgcaact acaacgccta ccttcgggcc agcgacgatc tgcctaagct gttcatcgag
     3121 tccgaccctg ggttcttttc caacgctatt gtcgagggag ctaagaagtt ccctaacacc
     3181 gagttcgtga aggtgaaggg cctccacttc ctccaggagg acgctccaga tgaaatgggt
     3241 aagtacatca agagcttcgt ggagcgcgtg ctgaagaacg agcagtaatc ggcgcgcctc
     3301 tagattaatt aaaagcttgg ccctattcta tagtgtcacc taaatgctag agctcgctga
     3361 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     3421 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     3481 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     3541 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     3601 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     3661 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3721 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3781 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3841 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3901 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3961 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     4021 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     4081 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4141 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4201 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4261 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4321 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     4381 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     4441 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4501 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4561 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4621 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4681 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4741 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4801 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4861 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4921 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4981 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     5041 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5101 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5161 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5221 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5281 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5341 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5401 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     5461 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5521 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5581 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5641 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5701 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5761 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5821 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5881 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5941 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6001 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6061 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6121 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6181 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6241 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6301 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6361 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6421 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6481 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6541 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6601 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6661 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6721 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6781 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6841 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6901 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6961 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7021 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7081 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7141 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7201 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7261 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7321 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7381 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7441 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7501 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7561 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7621 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7681 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7741 gcgcacattt ccccgaaaag tgccacctga cgtc
//
