LOCUS       sequence-469808-        6875 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of bovine arrestin-2 with C-terminal Flag tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6875)
  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 6875)
  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: p3A2-Flag
FEATURES             Location/Qualifiers
     source          1..6875
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(109..903)
                     /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(220..239)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     830..849
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(970..1299)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      984..1119
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1038..1057)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     1284..1304
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(1313..1741)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1423..1444)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1635..1654
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    1787..2011
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2000..2017
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        2037..2055
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             complement(2184..2207)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(3492..3510)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3601..3620)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3602..3620)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3665..3868)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3694..3714)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3869..4248
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4420..4439
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4514..4618
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4619..4687,4688..5479)
                     /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(4837..4856)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5650..6238
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6139..6158
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6392..6409
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6526..6547
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6562..6567,6568..6585,6586..6592)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6600..6616
                     /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     6605..6627
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6624..6640
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6624..6640
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    6689..6810
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6700..6719)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6754..6773
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 aaacgattcc gaagcccaac ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg
       61 gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc agaagaactc
      121 gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac
      181 gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc
      241 tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg
      301 gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc
      361 gccgtcgggc atgcgcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg
      421 ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc
      481 gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg
      541 ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag
      601 atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc
      661 gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc
      721 ctgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg
      781 cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata
      841 gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat
      901 catgcgaaac gatcctcatc ctgtctcttg atcagatccg aaaatggata tacaagctcc
      961 cgggagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag
     1021 ctcagaggca gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg
     1081 cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac
     1141 tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     1201 ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     1261 tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagaa ttaattcgcg
     1321 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     1381 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     1441 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     1501 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     1561 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     1621 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     1681 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     1741 tggggatacc ccctagagcc ccagctggtt ctttccgcct cagaagccat agagcccacc
     1801 gcatccccag catgcctgct attgtcttcc caatcctccc ccttgctgtc ctgccccacc
     1861 ccacccccca gaatagaatg acacctactc agacaatgcg atgcaatttc ctcattttat
     1921 taggaaagga cagtgggagt ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa
     1981 acaacagatg gctggcaact agaaggcaca gtcgaggctg atcagcgagc tctagcattt
     2041 aggtgacact atagaatagg gccccctggt agaaactgga agaacaaagg ggagggggag
     2101 ccaggacagg aagaagacga acaagcatcc tagtgcatga gagtggacct ggagctgccc
     2161 ggagggaggg cagccccagt ctacttatcg tcgtcatcct tgtaatctct gtcgttgagc
     2221 cgcggagagc cggtaccatc ctcctcttcc tccttgtcat ccttcatgcc ttttagtctc
     2281 tggcgggcaa agtcctcaaa cacaatgtcg tcatcgttgg tgtcaagttc tatgagattg
     2341 gtatctaccg gcgtctcgtg ctctggaact tcccggtgtg ggggttcctc tttgggcttg
     2401 gggtgcatta gggtgaaagg cagttccacg gccacatcac tggatgcaag atctcccaac
     2461 aggccgccac gagacaccac cagcttcact ttcactttgt aggaaacaat gatgcccagg
     2521 atctcccggt tggctccttc cctcaacagg gtgctggagg ccaggttcgt gtcctcgtgt
     2581 ttgagcttcc cgtccagggc gaggccccgc ttctctcgat tgttggccag gaagggggtc
     2641 agcgtgtaga ccttgcagaa cgtagagctg ggtgccactg tgtcatcagc ctcttccatg
     2701 gccacagggc acttgtactg ggctgtgttg aacagacaga tgtctgcata ctggcgcacc
     2761 gagatcttga tcttcttcac cgtcttgttg gtgttgttgg tgacatggac gttgacactg
     2821 atgggttctc cgtggtagta gatctccttg tccagggagg cctccagatg caagggcttg
     2881 tctgacatga ggaactgcct ggtggtctcg gccgtgggct gggggccagg cctctctggg
     2941 gcatactgaa ccttccggat gaccaggcgc acagaattcc gcttgtggat cttctcctcc
     3001 aggttctccg cacagaaggc tttcacttcg tagtccacac cgcaggcctt ccctgtatct
     3061 tcaggtcccg gctgcaaagt cacagagcat gggaggttcg gagggatctc aaaggtgaaa
     3121 gggtaggcat gctcgcccag cttcttgatg aggcgctcct gcagccgcgt caggggcttc
     3181 ttgtcctcag gggccggcgg gaaagactgc acgttggcca caaacaggtc cttgcgaaag
     3241 gtcaggccca ggacatccag gtcctcccgg ccgtagcgga aggcgcaggt cagcgtcaca
     3301 tagactctcc tctccttgag atactccgga tccacaagaa ccactccatc cacgggctcc
     3361 acgaggtcga tgtggtccac aaagtcccgc tttcccagat agacggtgag ctttccattg
     3421 gggctcgcct tcttgaacac ccgcgtccct ttgtcgccca tggtggttta aaaaaaaaag
     3481 aattcttgta ttctatagtg tcacctaaat cgcggccgcc agtgtgatgg atatctgcag
     3541 aattccagca cactggcggc cgttactagt ggatccgagc tcggtaccaa gcttgggtct
     3601 ccctatagtg agtcgtatta atttcgataa gccagtaagc agtgggttct ctagttagcc
     3661 agagagctct gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg
     3721 gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc
     3781 cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc
     3841 cattgatgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt
     3901 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt
     3961 taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca
     4021 agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg
     4081 ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc
     4141 caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta
     4201 atgaccccgt aattgattac tattaataac tagtcaataa tcaatgtcaa cgcgtatatc
     4261 tggcccgtac atcgcgaagc agcgcaaaac gcctaaccct aagcagattc ttcatgcaat
     4321 tgtcggtcaa gccttgcctt gttgtagctt aaattttgct cgcgcactac tcagcgacct
     4381 ccaacacaca agcagggagc agatactggc ttaactatgc ggcatcagag cagattgtac
     4441 tgagagtgca ccatagggga tcgggagatc tcccgatccg tcgacgtcag gtggcacttt
     4501 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4561 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     4621 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4681 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     4741 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     4801 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4861 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4921 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4981 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     5041 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     5101 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     5161 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     5221 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     5281 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5341 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5401 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5461 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     5521 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5581 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5641 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5701 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5761 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5821 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5881 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5941 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6001 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6061 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6121 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6181 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6241 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6301 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     6361 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     6421 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     6481 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     6541 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     6601 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagctcta
     6661 gctagaggtc gacggtatac agacatgata agatacattg atgagtttgg acaaaccaca
     6721 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     6781 gtaaccatta taagctgcaa taaacaagtt ggggtgggcg aagaactcca gcatgagatc
     6841 cccgcgctgg aggatcatcc agccggcgtc ccgga
//
