LOCUS       sequence-469811-        7369 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of bovine arrestin-3 (D291R).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7369)
  AUTHORS   Zheng C, Weinstein LD, Nguyen KK, Grewal A, Gurevich EV, Gurevich VV
  TITLE     GPCR Binding and JNK3 Activation by Arrestin-3 Have Different 
            Structural Requirements.
  JOURNAL   Cells. 2023 Jun 6;12(12):1563. doi: 10.3390/cells12121563.
  PUBMED    37371033
REFERENCE   2  (bases 1 to 7369)
  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: "Cells."; 
            date: "2023-06-6"; volume: "12(12):1563. doi"; pages: " 
            10.3390/cells12121563"
COMMENT     Sequence Label: pVEA3-DR
FEATURES             Location/Qualifiers
     source          1..7369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(6..28)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    17..33
                     /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(41..47,48..65,66..71))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    86..107
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(224..241)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(395..983)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(475..494)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1154..1945,1946..2014))
                     /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     1777..1796
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2015..2119)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2194..2213)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2385..2764
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2765..2968
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2919..2939
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3013..3032
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3013..3031
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(3066..3068,3069..3071,3072..3782)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3111..3132)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3372..3391)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3719..3740
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(5072..5090)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5073..5090)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5110..5127)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5116..5340
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5386..5814
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5473..5492)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5683..5704
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5823..5843)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5828..6157
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6008..6143
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6070..6089
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6224..7018
                     /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(6278..6297)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6888..6907
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7192..7313
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7229..7248)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7283..7302
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(join(7362..7369,1..9))
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(join(7362..7369,1..9))
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
       61 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
      121 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
      181 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
      241 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
      301 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
      361 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
      421 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
      481 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
      541 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
      601 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
      661 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
      721 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
      781 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
      841 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
      901 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
      961 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     1021 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     1081 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1141 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     1201 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     1261 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     1321 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     1381 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     1441 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     1501 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     1561 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     1621 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     1681 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     1741 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     1801 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     1861 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     1921 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     1981 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     2041 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     2101 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     2161 gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     2221 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
     2281 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
     2341 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
     2401 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     2461 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     2521 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     2581 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     2641 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     2701 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     2761 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     2821 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     2881 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     2941 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
     3001 gcttatcgaa attaatacga ctcactatag ggagacccaa gctggaattc gcgatcgcgg
     3061 ccacgatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     3121 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     3181 cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc gtgccctggc
     3241 ccaccctcgt gaccaccctg ggctacggcc tgcagtgctt cgcccgctac cccgaccaca
     3301 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     3361 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     3421 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     3481 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc gacaagcaga
     3541 agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc
     3601 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     3661 accactacct gagctaccag tccaaactga gcaaagaccc caacgagaag cgcgatcaca
     3721 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     3781 agtccggact taagtctcgg cgagcgctcg actccatggg ggagaaaccc gggaccaggg
     3841 tcttcaagaa atcgagtcct aactgcaagc tcaccgtgta cctaggcaag cgggactttg
     3901 tggaccacct ggacaaagtg gaccctgtag atggtgtggt gctagtggat ccggactact
     3961 tgaaggaccg caaagtgttt gtgaccctca cgtgcgcctt tcgctatggc cgcgaagacc
     4021 tggacgtcct gggcttgtcc ttccgcaaag acctgttcat cgccaactac caggccttcc
     4081 ctccaacacc taacccacct agaccaccta cgcgtctgca ggagcggctg ctgaggaagc
     4141 tgggccagca tgcccacccc ttttttttca caatacccca gaacctgccc tgctccgtca
     4201 ccctacagcc aggcccagag gatacgggga aggcttgcgg ggtcgacttt gagattcgag
     4261 ccttctgcgc caaatcacta gaagagaaaa gccacaaacg gaactccgta cgtctggtga
     4321 tccgaaaggt gcagtttgcc ccagagaaac cagggcccca gccttcagct gaaaccacac
     4381 gccacttcct catgtctgac cggtccctgc accttgaggc ttcactggac aaggagctgt
     4441 actaccacgg ggagcccctc aatgtcaatg tccacgtcac caataactcc accaagactg
     4501 tcaagaagat caaagtctct gtgagacagt acgccgacat ctgcctcttc agcaccgccc
     4561 agtacaagtg ccccgtggct caggtcgaac aagatgacca ggtgtcgccc agttccacgt
     4621 tctgtaaggt gtacaccata accccgctgc tcagcaacaa ccgggagaag cgcggcctcg
     4681 ctctacgtgg gaagctcaag cacgaggaca ccaacctggc ctcgagcacc attgtgaagg
     4741 agggtgccaa caaggaggtg ctggggatac tggtgtccta cagggtcaag gtgaagctgg
     4801 tggtgtcacg aggcggggat gtctcagtgg agctgccatt tgttctcatg caccccaagc
     4861 cccatgatca catcgccctt cccaggcccc agtcagctgt tcctgaaaca gatgcccccg
     4921 tggacaccaa cctcattgaa tttgaaacca actatgccac agacgacgac atcgtgttcg
     4981 aagactttgc ccggctgcgg ctcaaggggc tgaaggacga ggactatgac gaccagttct
     5041 gtatcgatta gggctggaag cttggcccta ttctatagtg tcacctaaat gctagagctc
     5101 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     5161 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     5221 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     5281 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     5341 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     5401 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     5461 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     5521 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     5581 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     5641 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     5701 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     5761 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     5821 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5881 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     5941 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     6001 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     6061 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     6121 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     6181 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     6241 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     6301 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     6361 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     6421 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     6481 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     6541 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     6601 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     6661 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     6721 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     6781 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     6841 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     6901 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     6961 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     7021 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     7081 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     7141 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     7201 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     7261 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     7321 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagc
//
