LOCUS       sequence-469809-        6100 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of bovine arrestin-3 with C-terminal HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6100)
  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 6100)
  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-HA
FEATURES             Location/Qualifiers
     source          1..6100
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3..25
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     22..38
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     22..38
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    87..208
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(98..117)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     152..171
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(382..1176)
                     /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(493..512)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1103..1122
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1243..1572)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1257..1392
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1311..1330)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     1557..1577
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(1586..2014)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1696..1717)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1908..1927
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2060..2284
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2273..2290
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        2310..2328
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2310..2327
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(2346..2372)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3099..3118)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3100..3118)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3163..3366)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3192..3212)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3367..3746
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3918..3937
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4012..4116
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4117..4185,4186..4977)
                     /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(4335..4354)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5148..5736
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5637..5656
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5890..5907
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6024..6045
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6060..6065,6066..6083,6084..6090)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    join(6098..6100,1..14)
                     /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)."
ORIGIN
        1 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagctctagc
       61 tagaggtcga cggtatacag acatgataag atacattgat gagtttggac aaaccacaac
      121 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
      181 aaccattata agctgcaata aacaagttgg ggtgggcgaa gaactccagc atgagatccc
      241 cgcgctggag gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat
      301 agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca
      361 tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg
      421 ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc
      481 aagctcttca gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc
      541 cagccggcca cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa
      601 gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg ggcatgcgcg ccttgagcct
      661 ggcgaacagt tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac
      721 aagaccggct tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa
      781 tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac
      841 tttctcggca ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag
      901 cagccagtcc cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt
      961 cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt tcattcaggg caccggacag
     1021 gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc
     1081 agagcagccg attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc
     1141 cggagaacct gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc
     1201 ttgatcagat ccgaaaatgg atatacaagc tcccgggagc tttttgcaaa agcctaggcc
     1261 tccaaaaaag cctcctcact acttctggaa tagctcagag gcagaggcgg cctcggcctc
     1321 tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt
     1381 taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat
     1441 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta
     1501 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc
     1561 taactgacac acattccaca gaattaattc gcgttaaatt tttgttaaat cagctcattt
     1621 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     1681 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     1741 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     1801 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     1861 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     1921 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     1981 cccgccgcgc ttaatgcgcc gctacagggc gcgtggggat accccctaga gccccagctg
     2041 gttctttccg cctcagaagc catagagccc accgcatccc cagcatgcct gctattgtct
     2101 tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga atgacaccta
     2161 ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg agtggcacct
     2221 tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc
     2281 acagtcgagg ctgatcagcg agctctagca tttaggtgac actatagaat agggccaagc
     2341 ttctacgcat agtctggaac atcgtaagga taatcgatac agaactggtc gtcatagtcc
     2401 tcgtccttca gccccttgag ccgcagccgg gcaaagtctt cgaacacgat gtcgtcgtct
     2461 gtggcatagt tggtttcaaa ttcaatgagg ttggtgtcca cgggggcatc tgtttcagga
     2521 acagctgact ggggcctggg aagggcgatg tgatcatggg gcttggggtg catgagaaca
     2581 aatggcagct ccactgagac atccccgcct cgtgacacca ccagcttcac cttgaccctg
     2641 taggacacca gtatccccag cacctccttg ttggcaccct ccttcacaat ggtgctcgag
     2701 gccaggttgg tgtcctcgtg cttgagcttc ccatctagag cgaggccgcg cttctcccgg
     2761 ttgttgctga gcagcggggt tatggtgtac accttacaga acgtggaact gggcgacacc
     2821 tggtcatctt gttcgacctg agccacgggg cacttgtact gggcggtgct gaagaggcag
     2881 atgtcggcgt actgtctcac agagactttg atcttcttga cagtcttggt ggagttattg
     2941 gtgacgtgga cattgacatt gaggggctcc ccgtggtagt acagctcctt gtccagtgaa
     3001 gcctcaaggt gcagggaccg gtcagacatg aggaagtggc gtgtggtttc agctgaaggc
     3061 tggggcccgg acatggtggt gaattccagc ttgggtctcc ctatagtgag tcgtattaat
     3121 ttcgataagc cagtaagcag tgggttctct agttagccag agagctctgc ttatatagac
     3181 ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt
     3241 tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag
     3301 acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg
     3361 catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca
     3421 taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg
     3481 gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata
     3541 ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat
     3601 tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag
     3661 ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta
     3721 ttaataacta gtcaataatc aatgtcaacg cgtatatctg gcccgtacat cgcgaagcag
     3781 cgcaaaacgc ctaaccctaa gcagattctt catgcaattg tcggtcaagc cttgccttgt
     3841 tgtagcttaa attttgctcg cgcactactc agcgacctcc aacacacaag cagggagcag
     3901 atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc ataggggatc
     3961 gggagatctc ccgatccgtc gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4021 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4081 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4141 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4201 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4261 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     4321 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     4381 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     4441 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     4501 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     4561 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     4621 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     4681 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     4741 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     4801 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     4861 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     4921 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     4981 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5041 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5101 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5161 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5221 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5281 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5341 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5401 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5461 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5521 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5581 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5641 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5701 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5761 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     5821 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     5881 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     5941 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     6001 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     6061 ttacacttta tgcttccggc tcgtatgttg tgtggaattg
//
