LOCUS       sequence-469807-        7369 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of bovine arrestin-3 (K11A, K12A, L49A, D51A, 
            R52A, L69A, Y239A, D241A, C252A, P253A, D260A, Q262A).
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: pVEA3-KNC
FEATURES             Location/Qualifiers
     source          1..7369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(18..39)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     367..386
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     626..647
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(726..745)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(727..745)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(790..993)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(819..839)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        994..1373
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1545..1564
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1639..1743
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1744..1812,1813..2604)
                     /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(1962..1981)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2775..3363
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3264..3283
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3517..3534
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3651..3672
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3687..3692,3693..3710,3711..3717)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3725..3741
                     /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     3730..3752
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3749..3765
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3749..3765
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3814..3935
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3825..3844)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3879..3898
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4109..4903)
                     /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(4220..4239)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4830..4849
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4970..5299)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4984..5119
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5038..5057)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     5284..5304
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(5313..5741)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5423..5444)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5635..5654
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    5787..6011
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6000..6017
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        6037..6055
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6037..6054
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(join(7345..7369,1..686,687..689,690..692))
                     /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"
ORIGIN
        1 gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg
       61 tctttgctca gtttggactg gtagctcagg tagtggttgt cgggcagcag cacggggccg
      121 tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgccgcc gtcctcgatg
      181 ttgtggcgga tcttgaagtt ggccttgatg ccgttcttct gcttgtcggc ggtgatatag
      241 acgttgtggc tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg
      301 aagtcgatgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg
      361 gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct
      421 tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcgggcgaag
      481 cactgcaggc cgtagcccag ggtggtcacg agggtgggcc agggcacggg cagcttgccg
      541 gtggtgcaga tcagcttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg
      601 gacacgctga acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc
      661 ccggtgaaca gctcctcgcc cttgctcacc atcgtggccg cgatcgcgaa ttccagcttg
      721 ggtctcccta tagtgagtcg tattaatttc gataagccag taagcagtgg gttctctagt
      781 tagccagaga gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt
      841 caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa
      901 actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc
      961 acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta
     1021 gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg
     1081 ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac
     1141 tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat
     1201 tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg
     1261 tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg
     1321 aactaatgac cccgtaattg attactatta ataactagtc aataatcaat gtcaacgcgt
     1381 atatctggcc cgtacatcgc gaagcagcgc aaaacgccta accctaagca gattcttcat
     1441 gcaattgtcg gtcaagcctt gccttgttgt agcttaaatt ttgctcgcgc actactcagc
     1501 gacctccaac acacaagcag ggagcagata ctggcttaac tatgcggcat cagagcagat
     1561 tgtactgaga gtgcaccata ggggatcggg agatctcccg atccgtcgac gtcaggtggc
     1621 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     1681 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     1741 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     1801 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     1861 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     1921 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     1981 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     2041 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     2101 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     2161 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     2221 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     2281 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     2341 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     2401 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     2461 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     2521 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     2581 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     2641 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     2701 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     2761 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2821 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2881 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     2941 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3001 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3061 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3121 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3181 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3241 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3301 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3361 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3421 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     3481 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     3541 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     3601 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     3661 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     3721 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     3781 ctctagctag aggtcgacgg tatacagaca tgataagata cattgatgag tttggacaaa
     3841 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     3901 tatttgtaac cattataagc tgcaataaac aagttggggt gggcgaagaa ctccagcatg
     3961 agatccccgc gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac
     4021 ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg
     4081 tcggtcattt cgaaccccag agtcccgctc agaagaactc gtcaagaagg cgatagaagg
     4141 cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt
     4201 cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg
     4261 ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat
     4321 tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgcgcgcct
     4381 tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct
     4441 gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt
     4501 ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga
     4561 tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc
     4621 ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa
     4681 cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ctgcagttca ttcagggcac
     4741 cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg
     4801 cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc
     4861 aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc
     4921 ctgtctcttg atcagatccg aaaatggata tacaagctcc cgggagcttt ttgcaaaagc
     4981 ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggca gaggcggcct
     5041 cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg
     5101 gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga
     5161 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg
     5221 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
     5281 cacaccctaa ctgacacaca ttccacagaa ttaattcgcg ttaaattttt gttaaatcag
     5341 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     5401 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     5461 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     5521 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     5581 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     5641 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     5701 caccacaccc gccgcgctta atgcgccgct acagggcgcg tggggatacc ccctagagcc
     5761 ccagctggtt ctttccgcct cagaagccat agagcccacc gcatccccag catgcctgct
     5821 attgtcttcc caatcctccc ccttgctgtc ctgccccacc ccacccccca gaatagaatg
     5881 acacctactc agacaatgcg atgcaatttc ctcattttat taggaaagga cagtgggagt
     5941 ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa acaacagatg gctggcaact
     6001 agaaggcaca gtcgaggctg atcagcgagc tctagcattt aggtgacact atagaatagg
     6061 gccaagcttc cagccctaat cgatacagaa ctggtcgtca tagtcctcgt ccttcagccc
     6121 cttgagccgc agccgggcaa agtcttcgaa cacgatgtcg tcgtctgtgg catagttggt
     6181 ttcaaattca atgaggttgg tgtccacggg ggcatctgtt tcaggaacag ctgactgggg
     6241 cctgggaagg gcgatgtgat catggggctt ggggtgcatg agaacaaatg gcagctccac
     6301 tgagacatcc ccgcctcgtg acaccaccag cttcaccttg accctgtagg acaccagtat
     6361 ccccagcacc tccttgttgg caccctcctt cacaatggtg ctcgaggcca ggttggtgtc
     6421 ctcgtgcttg agcttcccat ctagagcgag gccgcgcttc tcccggttgt tgctgagcag
     6481 cggggttatg gtgtacacct tacagaacgt ggaactgggc gacacagcgt ctgcttgttc
     6541 gacctgagcc acggccgcct tgtactgggc ggtgctgaag aggcagatag cggctgcctg
     6601 tctcacagag actttgatct tcttgacagt cttggtggag ttattggtga cgtggacatt
     6661 gacattgagg ggctccccgt ggtagtacag ctccttgtcc agtgaagcct caaggtgcag
     6721 ggaccggtca gacatgagga agtggcgtgt ggtttcagct gaaggctggg gccctggttt
     6781 ctctggggca aactgcacct ttcggatcac cagacgtacg gagttccgtt tgtggctttt
     6841 ctcttctagt gatttggcgc agaaggctcg aatctcaaag tcgaccccgc aagccttccc
     6901 cgtatcctct gggcctggct gtagggtgac ggagcagggc aggttctggg gtattgtgaa
     6961 aaaaaagggg tgggcatgct ggcccagctt cctcagcagc cgctcctgca gacgcgtagg
     7021 tggtctaggt gggttaggtg ttggagggaa ggcctggtag ttggcgatga acaggtcttt
     7081 gcggaaggac aagcccagga cgtcagcgtc ttcgcggcca tagcgaaagg cgcacgtgag
     7141 ggtcacaaac actttcgcgg ccttagcgta gtccggatcc actagcacca caccatctac
     7201 agggtccact ttgtccaggt ggtccacaaa gtcccgcttg cctaggtaca cggtgagctt
     7261 gcagttagga ctcgatgccg cgaagaccct ggtcccgggt ttctccccca tggagtcgag
     7321 cgctcgccga gacttaagtc cggacttgta cagctcgtcc atgccgaga
//
