LOCUS       sequence-469806-        6876 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of bovine arrestin-3 with C-terminal Flag tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6876)
  AUTHORS   Ahmed MR, Zhan X, Song X, Kook S, Gurevich VV, Gurevich EV
  TITLE     Ubiquitin ligase parkin promotes Mdm2-arrestin interaction but 
            inhibits arrestin ubiquitination.
  JOURNAL   Biochemistry. 2011 May 10;50(18):3749-63. doi: 10.1021/bi200175q. 
            Epub 2011 Apr 12.
  PUBMED    21466165
REFERENCE   2  (bases 1 to 6876)
  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: "10.1021/bi200175q";
            journalName: "Biochemistry"; date: "2011-05-10- 10"; volume: "50"; 
            issue: "18"; pages: "3749-63"
COMMENT     Sequence Label: p3A3-Flag
FEATURES             Location/Qualifiers
     source          1..6876
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      507..1095
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     996..1015
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1249..1266
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1383..1404
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1419..1424,1425..1442,1443..1449)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1457..1473
                     /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     1462..1484
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1481..1497
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1481..1497
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1546..1667
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1557..1576)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1611..1630
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1841..2635)
                     /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(1952..1971)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2562..2581
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2702..3031)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2716..2851
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2770..2789)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     3016..3036
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(3045..3473)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3155..3176)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3367..3386
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3519..3743
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3732..3749
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        3769..3787
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             complement(3941..3964)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(5225..5243)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5334..5353)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5335..5353)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(5398..5601)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5427..5447)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5602..5981
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     6153..6172
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6247..6351
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6352..6420,6421..6876,1..336)
                     /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(6570..6589)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
       61 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
      121 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
      181 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
      241 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
      301 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
      361 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
      421 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
      481 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
      541 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
      601 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
      661 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
      721 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
      781 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
      841 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
      901 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
      961 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     1021 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     1081 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     1141 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     1201 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     1261 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     1321 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     1381 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     1441 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     1501 gattacgcca agctctagct agaggtcgac ggtatacaga catgataaga tacattgatg
     1561 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     1621 atgctattgc tttatttgta accattataa gctgcaataa acaagttggg gtgggcgaag
     1681 aactccagca tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacgatt
     1741 ccgaagccca acctttcata gaaggcggcg gtggaatcga aatctcgtga tggcaggttg
     1801 ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa
     1861 ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc
     1921 ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct
     1981 gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt
     2041 ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg
     2101 gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt
     2161 ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat
     2221 gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg
     2281 catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc
     2341 ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag
     2401 ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcctgcagtt
     2461 cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca
     2521 gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata
     2581 gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa
     2641 acgatcctca tcctgtctct tgatcagatc cgaaaatgga tatacaagct cccgggagct
     2701 ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg
     2761 cagaggcggc ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat
     2821 gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg
     2881 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
     2941 cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
     3001 ctggggactt tccacaccct aactgacaca cattccacag aattaattcg cgttaaattt
     3061 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     3121 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     3181 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     3241 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     3301 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     3361 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     3421 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtggggata
     3481 ccccctagag ccccagctgg ttctttccgc ctcagaagcc atagagccca ccgcatcccc
     3541 agcatgcctg ctattgtctt cccaatcctc ccccttgctg tcctgcccca ccccaccccc
     3601 cagaatagaa tgacacctac tcagacaatg cgatgcaatt tcctcatttt attaggaaag
     3661 gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga
     3721 tggctggcaa ctagaaggca cagtcgaggc tgatcagcga gctctagcat ttaggtgaca
     3781 ctatagaata gggccccctg gtagaaactg gaagaacaaa ggggaggggg agccaggaca
     3841 ggaagaagac gaacaagcat cctagtgcat gagagtggac ctggagctgc ccggagggag
     3901 ggcagcccca gtctatctgt cgttgagccg ctctagacta cttatcgtcg tcatccttgt
     3961 agtcgacgca gaactggtcg tcatagtcct cgtccttcag ccccttgagc cgcagccggg
     4021 caaagtcctc aaacacgatg tcgtcgtctg tggcatagtt ggtttcgaat tcaatgaggt
     4081 tggtgtccac gggggcatct gtttcaggaa cagctgactg gggcctggga agggcgatgt
     4141 gatcatgggg cttggggtgc atgagaacaa atggcagctc cactgagacg tccccgcctc
     4201 gagacaccac cagcttcacc ttgaccctgt aggacaccag gatccccagc acctccttgt
     4261 tggcaccctc cttcacaatg gtgctggacg ccaggttggt gtcctcgtgc ttgagcttcc
     4321 catccagagc gaggccgcgc ttctcccggt tgttgctgag cagcggggtt atggtgtaca
     4381 ccttacagaa cgtggaactg ggcgacacct ggtcatcttg ttcgacctga gccacggggc
     4441 acttgtactg ggcggtgctg aagaggcaga tgtcggcgta ctgtctcaca gagactttga
     4501 tcttcttgac agtcttggtg gagttattgg tgacgtggac attgacattg aggggctccc
     4561 catggtagta cagctccttg tccagtgaag cctcaaggtg cagggaccgg tcagacatga
     4621 ggaagtggcg tgtggtttca gctgaaggct gggggccggg tttctctggg gcaaactgca
     4681 cctttcggat caccagacgc acagagttcc gtttgtggct tttctcttct agtgatttgg
     4741 cgcagaaggc tcgaatctca aagtctaccc cgcaggcctt ccccgtatcc tctgggcctg
     4801 gctgcagggt gacggagcag ggcaggttct ggggtattgt gaaaaaaaag gggtgggcat
     4861 gctggcccag cttcctcagc agccgctcct gcaggcgggt ggggggccgg ggtgggttgg
     4921 gtgttggggg gaaggcctgg tagttggcga tgaacaggtc tttgcggaag gacaagccca
     4981 gcacgtccag gtcttcgcgg ccatagcgaa aggcgcaggt gagggtcaca aacactttgc
     5041 ggtccttcaa gtagtccggg tccactagca ccacaccatc tacaggatcc actttgtcca
     5101 ggtggtccac aaagtcccgc ttgcccaggt acacggtgag cttgcagtta ggactcgatt
     5161 tcttgaagac cctggtcccg ggtttctccc ccatggtggt ttaaaaaaaa aagaattctt
     5221 gtattctata gtgtcaccta aatcgcggcc gccagtgtga tggatatctg cagaattcca
     5281 gcacactggc ggccgttact agtggatccg agctcggtac caagcttggg tctccctata
     5341 gtgagtcgta ttaatttcga taagccagta agcagtgggt tctctagtta gccagagagc
     5401 tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca atggggcgga
     5461 gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac
     5521 gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat
     5581 gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga tgtactgcca
     5641 agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc
     5701 attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc
     5761 agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat
     5821 gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg
     5881 gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc
     5941 cgtaattgat tactattaat aactagtcaa taatcaatgt caacgcgtat atctggcccg
     6001 tacatcgcga agcagcgcaa aacgcctaac cctaagcaga ttcttcatgc aattgtcggt
     6061 caagccttgc cttgttgtag cttaaatttt gctcgcgcac tactcagcga cctccaacac
     6121 acaagcaggg agcagatact ggcttaacta tgcggcatca gagcagattg tactgagagt
     6181 gcaccatagg ggatcgggag atctcccgat ccgtcgacgt caggtggcac ttttcgggga
     6241 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     6301 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     6361 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     6421 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     6481 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     6541 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     6601 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     6661 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     6721 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     6781 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     6841 gaaccggagc tgaatgaagc cataccaaac gacgag
//
