LOCUS       sequence-470240-        6670 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of bovine arrestin-2 with C-terminal HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6670)
  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 6670)
  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-HA
FEATURES             Location/Qualifiers
     source          1..6670
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             660..686
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(704..722)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(705..722)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(742..759)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    748..972
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1018..1446
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1105..1124)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1315..1336
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1455..1475)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1460..1789
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1640..1775
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1702..1721
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1856..2650
                     /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(1910..1929)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2520..2539
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2824..2945
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2861..2880)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2915..2934
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2994..3010)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2994..3010)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3007..3029)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3018..3034
                     /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)."
     promoter        complement(join(3042..3048,3049..3066,3067..3072))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3087..3108
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3225..3242)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3396..3984)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3476..3495)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4155..4946,4947..5015))
                     /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     4778..4797
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5016..5120)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5195..5214)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5386..5765
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5766..5969
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5920..5940
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6014..6033
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6014..6032
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cctccctgga caaggagatc tactaccacg gagaacccat cagtgtcaac gtccatgtca
       61 ccaacaacac caacaagacg gtgaagaaga tcaagatctc ggtgcgccag tatgcagaca
      121 tctgtctgtt caacacagcc cagtacaagt gccctgtggc catggaagag gctgatgaca
      181 cagtggcacc cagctctacg ttctgcaagg tctacacgct gacccccttc ctggccaaca
      241 atcgagagaa gcggggcctc gctctagacg ggaagctcaa acacgaggac acgaacctgg
      301 cctcgagcac cctgttgagg gaaggagcca accgggaaat cctgggcatc attgtttcct
      361 acaaagtgaa agtgaagctg gtggtgtctc gtggcggcct gttgggagat cttgcatcca
      421 gtgatgtggc cgtggaactg cctttcaccc taatgcaccc caagcccaaa gaggaacccc
      481 cacaccggga agttccagag cacgagacgc cggtagatac caatctcata gaacttgaca
      541 ccaacgatga cgacattgtg ttcgaagact ttgcccgcca gagactaaaa ggcatgaagg
      601 atgacaagga ggaagaggag gatggtaccg gctctccgcg gctcaacgac agaatcgatt
      661 atccttacga tgttccagac tatgcgtaga agcttggccc tattctatag tgtcacctaa
      721 atgctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
      781 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
      841 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
      901 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
      961 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     1021 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     1081 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     1141 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     1201 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     1261 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     1321 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     1381 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     1441 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     1501 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     1561 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     1621 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     1681 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     1741 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     1801 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     1861 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     1921 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     1981 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     2041 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     2101 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     2161 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     2221 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     2281 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     2341 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     2401 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     2461 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     2521 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     2581 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     2641 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     2701 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     2761 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     2821 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     2881 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     2941 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     3001 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     3061 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     3121 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3181 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3241 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3301 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3361 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3421 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3481 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3541 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3601 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3661 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3721 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3781 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3841 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3901 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3961 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4021 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4081 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4141 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4201 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4261 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4321 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4381 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4441 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4501 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4561 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4621 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4681 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4741 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     4801 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4861 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4921 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4981 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5041 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5101 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     5161 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     5221 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     5281 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
     5341 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga
     5401 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     5461 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     5521 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     5581 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     5641 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     5701 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     5761 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     5821 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     5881 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     5941 tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact
     6001 ggcttatcga aattaatacg actcactata gggagaccca agctggaatt cttttttttt
     6061 taaaccacca tgggcgacaa agggacgcgg gtgttcaaga aggcgagccc caatggaaag
     6121 ctcaccgtct acctaggaaa gcgggacttt gtggaccaca tcgacctcgt ggagcccgtg
     6181 gatggagtgg ttcttgtgga tccggagtat ctcaaggaga ggagagtcta tgtgacgctc
     6241 acgtgcgcct tccgctacgg acgggaggac ctggacgtcc tgggcctgac ctttcgcaag
     6301 gacctgtttg tggccaacgt gcagtctttc ccgccggccc ctgaggacaa gaagcccctg
     6361 acgcgtctgc aggagcgcct catcaagaag ctgggcgagc atgcctaccc tttcaccttt
     6421 gagatccctc cgaacctgcc atgctctgtg actttgcagc cgggacctga agatacaggg
     6481 aaggcttgcg gtgtcgacta cgaagtgaaa gccttctgtg cggagaacct ggaggagaag
     6541 atccacaagc ggaactccgt acgcctggtc atccggaagg ttcagtatgc cccagagagg
     6601 cccgggcccc agcccacggc cgagaccacc aggcagttcc tcatgtcaga caagcccttg
     6661 catctggagg
//
