LOCUS       sequence-471661-        7513 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human angiotensin II type 1 receptor; 
            N-terminal HA tag, C-terminal Rluc8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7513)
  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 7513)
  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: p3HAT1Luc1
FEATURES             Location/Qualifiers
     source          1..7513
                     /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             2092..3027
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNATSSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGAALAFHYAYEHQDRIKAIVHMESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETVLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKLFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFLQEDAPDEMGKYIKSFVERVLKNEQ"
     promoter        complement(3066..3084)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3067..3084)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3104..3121)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3110..3334
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3380..3808
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3467..3486)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3677..3698
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3817..3837)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3822..4151
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4002..4137
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4064..4083
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4218..5012
                     /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(4272..4291)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4882..4901
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5186..5307
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5223..5242)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5277..5296
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5356..5372)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5356..5372)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5369..5391)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5380..5396
                     /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(5404..5410,5411..5428,5429..5434))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5449..5470
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5587..5604)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5758..6346)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5838..5857)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6517..7308,7309..7377))
                     /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     7140..7159
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7378..7482)
                     /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 ctgatattct caactcttct
     1021 actgaagatg gtattaaaag aatccaagat gattgtccca aagctggaag gcataattac
     1081 atatttgtca tgattcctac tttatacagt atcatctttg tggtgggaat atttggaaac
     1141 agcttggtgg tgatagtcat ttacttttat atgaagctga agactgtggc cagtgttttt
     1201 cttttgaatt tagcactggc tgacttatgc tttttactga ctttgccact atgggctgtc
     1261 tacacagcta tggaataccg ctggcccttt ggcaattacc tatgtaagat tgcttcagcc
     1321 agcgtcagtt tcaacctgta cgctagtgtg tttctactca cgtgtctcag cattgatcga
     1381 tacctggcta ttgttcaccc aatgaagtcc cgccttcgac gcacaatgct tgtagccaaa
     1441 gtcacctgca tcatcatttg gctgctggca ggcttggcca gtttgccagc tataatccat
     1501 cgaaatgtat ttttcattga gaacaccaat attacagttt gtgctttcca ttatgagtcc
     1561 caaaattcaa cccttccgat agggctgggc ctgaccaaaa atatactggg tttcctgttt
     1621 ccttttctga tcattcttac aagttatact cttatttgga aggccctaaa gaaggcttat
     1681 gaaattcaga agaacaaacc aagaaatgat gatattttta agataattat ggcaattgtg
     1741 cttttctttt tcttttcctg gattccccac caaatattca cttttctgga tgtattgatt
     1801 caactaggca tcatacgtga ctgtagaatt gcagatattg tggacacggc catgcctatc
     1861 accatttgta tagcttattt taacaattgc ctgaatcctc ttttttatgg ctttctgggg
     1921 aaaaaattta aaagatattt tctccagctt ctaaaatata ttcccccaaa agccaaatcc
     1981 cactcaaacc tttcaacaaa aatgagcacg ctttcctacc gcccctcaga taatgtaagc
     2041 tcatccacca agaagcctgc accatgtttt gaggttgagg cacctgcagg catggcttcc
     2101 aaggtgtacg accccgagca acgcaaacgc atgatcactg ggcctcagtg gtgggctcgc
     2161 tgcaagcaaa tgaacgtgct ggactccttc atcaactact atgattccga gaagcacgcc
     2221 gagaacgccg tgatttttct gcatggtaac gctacctcca gctacctgtg gaggcacgtc
     2281 gtgcctcaca tcgagcccgt ggctagatgc atcatccctg atctgatcgg aatgggtaag
     2341 tccggcaaga gcgggaatgg ctcatatcgc ctcctggatc actacaagta cctcaccgct
     2401 tggttcgagc tgctgaacct tccaaagaaa atcatctttg tgggccacga ctggggggct
     2461 gctctggcct ttcactacgc ctacgagcac caagacagga tcaaggccat cgtccatatg
     2521 gagagtgtcg tggacgtgat cgagtcctgg gacgagtggc ctgacatcga ggaggatatc
     2581 gccctgatca agagcgaaga gggcgagaaa atggtgcttg agaataactt cttcgtcgag
     2641 accgtgctcc caagcaagat catgcggaaa ctggagcctg aggagttcgc tgcctacctg
     2701 gagccattca aggagaaggg cgaggttaga cggcctaccc tctcctggcc tcgcgagatc
     2761 cctctcgtta agggaggcaa gcccgacgtc gtccagattg tccgcaacta caacgcctac
     2821 cttcgggcca gcgacgatct gcctaagctg ttcatcgagt ccgaccctgg gttcttttcc
     2881 aacgctattg tcgagggagc taagaagttc cctaacaccg agttcgtgaa ggtgaagggc
     2941 ctccacttcc tccaggagga cgctccagat gaaatgggta agtacatcaa gagcttcgtg
     3001 gagcgcgtgc tgaagaacga gcagtaatcg gcgcgcctct agattaatta aaagcttggc
     3061 cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct
     3121 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     3181 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3241 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3301 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     3361 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3421 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3481 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3541 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3601 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3661 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3721 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     3781 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     3841 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3901 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3961 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4021 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4081 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4141 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     4201 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4261 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4321 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4381 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4441 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4501 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4561 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4621 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4681 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4741 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4801 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4861 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4921 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4981 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     5041 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5101 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5161 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     5221 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5281 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5341 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5401 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5461 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5521 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5581 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5641 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5701 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5761 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5821 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5881 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5941 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6001 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6061 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6121 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6181 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6241 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6301 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6361 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6421 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6481 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6541 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6601 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6661 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6721 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6781 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6841 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6901 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6961 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7021 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7081 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7141 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7201 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7261 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7321 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7381 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7441 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7501 gccacctgac gtc
//
