LOCUS       sequence-469804-        7369 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of bovine arrestin-3 (I385A,V386A,F387A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7369)
  AUTHORS   Breitman M, Kook S, Gimenez LE, Lizama BN, Palazzo MC, Gurevich EV, 
            Gurevich VV
  TITLE     Silent scaffolds: inhibition OF c-Jun N-terminal kinase 3 activity 
            in cell by dominant-negative arrestin-3 mutant.
  JOURNAL   J Biol Chem. 2012 Jun 1;287(23):19653-64. doi: 
            10.1074/jbc.M112.358192. Epub 2012 Apr 20.
  PUBMED    22523077
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"; doi: 
            "10.1074/jbc.M112.358192"; journalName: "J Biol Chem"; date: 
            "2012-06-1- 1"; volume: "287"; issue: "23"; pages: "19653-64"
COMMENT     Sequence Label: p3VEA3-3A
FEATURES             Location/Qualifiers
     source          1..7369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      754..1342
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1243..1262
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1496..1513
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1630..1651
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1666..1671,1672..1689,1690..1696)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1704..1720
                     /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     1709..1731
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1728..1744
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1728..1744
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1793..1914
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1804..1823)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1858..1877
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2088..2882)
                     /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(2199..2218)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2809..2828
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2949..3278)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2963..3098
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3017..3036)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     3263..3283
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(3292..3720)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3402..3423)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3614..3633
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3766..3990
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3979..3996
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        4016..4034
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4016..4033
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(join(5324..6034,6035..6037,6038..6040))
                     /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(5366..5387)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5715..5734
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5974..5995
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6074..6093)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6075..6093)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(6138..6341)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6167..6187)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6342..6721
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     6893..6912
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6987..7091
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7092..7160,7161..7369,1..583)
                     /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     complement(7310..7329)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
       61 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
      121 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
      181 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
      241 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
      301 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
      361 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
      421 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
      481 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
      541 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
      601 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
      661 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
      721 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
      781 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
      841 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
      901 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
      961 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     1021 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     1081 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     1141 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     1201 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     1261 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     1321 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     1381 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     1441 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     1501 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     1561 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     1621 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     1681 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     1741 tgaccatgat tacgccaagc tctagctaga ggtcgacggt atacagacat gataagatac
     1801 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     1861 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttggggtg
     1921 ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa
     1981 aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg
     2041 caggttgggc gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg
     2101 tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg
     2161 aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct
     2221 atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg
     2281 ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg
     2341 ccgtcgggca tgcgcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc
     2401 tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg
     2461 atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc
     2521 cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga
     2581 tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg
     2641 agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtcc
     2701 tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc
     2761 gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag
     2821 ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc
     2881 atgcgaaacg atcctcatcc tgtctcttga tcagatccga aaatggatat acaagctccc
     2941 gggagctttt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt ctggaatagc
     3001 tcagaggcag aggcggcctc ggcctctgca taaataaaaa aaattagtca gccatggggc
     3061 ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag gggcgggact
     3121 atggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg
     3181 gactttccac acctggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct
     3241 ggggagcctg gggactttcc acaccctaac tgacacacat tccacagaat taattcgcgt
     3301 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     3361 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     3421 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     3481 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     3541 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     3601 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     3661 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     3721 ggggataccc cctagagccc cagctggttc tttccgcctc agaagccata gagcccaccg
     3781 catccccagc atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc tgccccaccc
     3841 caccccccag aatagaatga cacctactca gacaatgcga tgcaatttcc tcattttatt
     3901 aggaaaggac agtgggagtg gcaccttcca gggtcaagga aggcacgggg gaggggcaaa
     3961 caacagatgg ctggcaacta gaaggcacag tcgaggctga tcagcgagct ctagcattta
     4021 ggtgacacta tagaataggg ccaagcttcc agccctaatc gatacagaac tggtcgtcat
     4081 agtcctcgtc cttcagcccc ttgagccgca gccgggcaaa gtcttcagcc gcggcgtcgt
     4141 cgtctgtggc atagttggtt tcaaattcaa tgaggttggt gtccacgggg gcatctgttt
     4201 caggaacagc tgactggggc ctgggaaggg cgatgtgatc atggggcttg gggtgcatga
     4261 gaacaaatgg cagctccact gagacatccc cgcctcgtga caccaccagc ttcaccttga
     4321 ccctgtagga caccagtatc cccagcacct ccttgttggc accctccttc acaatggtgc
     4381 tcgaggccag gttggtgtcc tcgtgcttga gcttcccatc tagagcgagg ccgcgcttct
     4441 cccggttgtt gctgagcagc ggggttatgg tgtacacctt acagaacgtg gaactgggcg
     4501 acacctggtc atcttgttcg acctgagcca cggggcactt gtactgggcg gtgctgaaga
     4561 ggcagatgtc ggcgtactgt ctcacagaga ctttgatctt cttgacagtc ttggtggagt
     4621 tattggtgac gtggacattg acattgaggg gctccccgtg gtagtacagc tccttgtcca
     4681 gtgaagcctc aaggtgcagg gaccggtcag acatgaggaa gtggcgtgtg gtttcagctg
     4741 aaggctgggg ccctggtttc tctggggcaa actgcacctt tcggatcacc agacgtacgg
     4801 agttccgttt gtggcttttc tcttctagtg atttggcgca gaaggctcga atctcaaagt
     4861 cgaccccgca agccttcccc gtatcctctg ggcctggctg tagggtgacg gagcagggca
     4921 ggttctgggg tattgtgaaa aaaaaggggt gggcatgctg gcccagcttc ctcagcagcc
     4981 gctcctgcag acgcgtaggt ggtctaggtg ggttaggtgt tggagggaag gcctggtagt
     5041 tggcgatgaa caggtctttg cggaaggaca agcccaggac gtccaggtct tcgcggccat
     5101 agcgaaaggc gcacgtgagg gtcacaaaca ctttgcggtc cttcaagtag tccggatcca
     5161 ctagcaccac accatctaca gggtccactt tgtccaggtg gtccacaaag tcccgcttgc
     5221 ctaggtacac ggtgagcttg cagttaggac tcgatttctt gaagaccctg gtcccgggtt
     5281 tctcccccat ggagtcgagc gctcgccgag acttaagtcc ggacttgtac agctcgtcca
     5341 tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct
     5401 cgttggggtc tttgctcagt ttggactggt agctcaggta gtggttgtcg ggcagcagca
     5461 cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgccgccgt
     5521 cctcgatgtt gtggcggatc ttgaagttgg ccttgatgcc gttcttctgc ttgtcggcgg
     5581 tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt
     5641 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
     5701 tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga
     5761 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
     5821 gggcgaagca ctgcaggccg tagcccaggg tggtcacgag ggtgggccag ggcacgggca
     5881 gcttgccggt ggtgcagatc agcttcaggg tcagcttgcc gtaggtggca tcgccctcgc
     5941 cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg
     6001 gcaccacccc ggtgaacagc tcctcgccct tgctcaccat cgtggccgcg atcgcgaatt
     6061 ccagcttggg tctccctata gtgagtcgta ttaatttcga taagccagta agcagtgggt
     6121 tctctagtta gccagagagc tctgcttata tagacctccc accgtacacg cctaccgccc
     6181 atttgcgtca atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc
     6241 caaaacaaac tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac
     6301 cgctatccac gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact
     6361 aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc
     6421 caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact
     6481 tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa
     6541 gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg
     6601 ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat
     6661 gggctatgaa ctaatgaccc cgtaattgat tactattaat aactagtcaa taatcaatgt
     6721 caacgcgtat atctggcccg tacatcgcga agcagcgcaa aacgcctaac cctaagcaga
     6781 ttcttcatgc aattgtcggt caagccttgc cttgttgtag cttaaatttt gctcgcgcac
     6841 tactcagcga cctccaacac acaagcaggg agcagatact ggcttaacta tgcggcatca
     6901 gagcagattg tactgagagt gcaccatagg ggatcgggag atctcccgat ccgtcgacgt
     6961 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     7021 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     7081 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     7141 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     7201 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     7261 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     7321 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcat
//
