LOCUS       sequence-473908-        7471 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mutant bovine arrestin-2 long isoform with 
            Venus N-terminal fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7471)
  AUTHORS   Gimenez LE, Kook S, Vishnivetskiy SA, Ahmed MR, Gurevich EV, 
            Gurevich VV
  TITLE     Role of receptor-attached phosphates in binding of visual and 
            non-visual arrestins to G protein-coupled receptors.
  JOURNAL   J Biol Chem. 2012 Mar 16;287(12):9028-40. doi: 
            10.1074/jbc.M111.311803. Epub 2012 Jan 24.
  PUBMED    22275358
REFERENCE   2  (bases 1 to 7471)
  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.M111.311803"; journalName: "J Biol Chem"; date: 
            "2012-03-16- 16"; volume: "287"; issue: "12"; pages: "9028-40"
COMMENT     Sequence Label: pVeBtA2-3A
FEATURES             Location/Qualifiers
     source          1..7471
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(916..918,919..921,922..1632)
                     /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"
     primer_bind     complement(961..982)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1222..1241)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1569..1590
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(3024..3042)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3025..3042)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3062..3079)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3068..3292
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3338..3766
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3425..3444)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3635..3656
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3775..3795)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3780..4109
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3960..4095
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4022..4041
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4176..4970
                     /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(4230..4249)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4840..4859
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5144..5265
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5181..5200)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5235..5254
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5314..5330)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5314..5330)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5327..5349)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5338..5354
                     /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(5362..5368,5369..5386,5387..5392))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5407..5428
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5545..5562)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5716..6304)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5796..5815)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6475..7266,7267..7335))
                     /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     7098..7117
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7336..7440)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggaattc
      901 gcgatcgcgg ccacgatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
      961 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     1021 ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc
     1081 gtgccctggc ccaccctcgt gaccaccctg ggctacggcc tgcagtgctt cgcccgctac
     1141 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1201 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1261 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1321 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc
     1381 gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc
     1441 ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1501 ctgcccgaca accactacct gagctaccag tccaaactga gcaaagaccc caacgagaag
     1561 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1621 gagctgtaca agtccggact taagtctcgg cgagcgctcg actccatggg cgacaaaggg
     1681 acgcgggtgt tcaagaaggc gagccccaat ggaaagctca ccgtctacct aggaaagcgg
     1741 gactttgtgg accacatcga cctcgtggag cccgtggatg gagtggttct tgtggatccg
     1801 gagtatctca aggagaggag agtctatgtg acgctcacgt gcgccttccg ctacggacgg
     1861 gaggacctgg acgtcctggg cctgaccttt cgcaaggacc tgtttgtggc caacgtgcag
     1921 tctttcccgc cggcccctga ggacaagaag cccctgacgc gtctgcagga gcgcctcatc
     1981 aagaagctgg gcgagcatgc ctaccctttc acctttgaga tccctccgaa cctgccatgc
     2041 tctgtgactt tgcagccggg acctgaagat acagggaagg cttgcggtgt cgactacgaa
     2101 gtgaaagcct tctgtgcgga gaacctggag gagaagatcc acaagcggaa ctccgtacgc
     2161 ctggtcatcc ggaaggttca gtatgcccca gagaggcccg ggccccagcc cacggccgag
     2221 accaccaggc agttcctcat gtcagacaag cccttgcatc tggaggcctc cctggacaag
     2281 gagatctact accacggaga acccatcagt gtcaacgtcc atgtcaccaa caacaccaac
     2341 aagacggtga agaagatcaa gatctcggtg cgccagtatg cagacatctg tctgttcaac
     2401 acagcccagt acaagtgccc tgtggccatg gaagaggctg atgacacagt ggcacccagc
     2461 tctacgttct gcaaggtcta cacgctgacc cccttcctgg ccaacaatcg agagaagcgg
     2521 ggcctcgccc tggacgggaa gctcaaacac gaggacacga acctggcctc cagcaccctg
     2581 ttgagggaag gagccaaccg ggagatcctg ggcatcattg tttcctacaa agtgaaagtg
     2641 aagctggtgg tgtctcgtgg cggcctgttg ggagatcttg catccagtga tgtggccgtg
     2701 gaactgcctt tcaccctaat gcaccccaag cccaaagagg aacccccaca ccgggaagtt
     2761 ccagagcacg agacgccggt agataccaat ctcatagaac ttgacaccaa cgatgacgac
     2821 gctgcggctg aggactttgc ccgccagaga ctaaaaggca tgaaggatga caaggaggaa
     2881 gaggaggatg gtaccggctc tccgcggctc aacgacagaa tcgattagcg gccgctgccc
     2941 tccctccggg cagctccagg tccactctca tgcactagga tgcttgttcg tcttcttcct
     3001 gtcctggcta agcttggccc tattctatag tgtcacctaa atgctagagc tcgctgatca
     3061 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     3121 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3181 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     3241 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     3301 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     3361 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3421 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3481 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3541 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3601 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3661 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3721 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     3781 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3841 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3901 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3961 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4021 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     4081 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     4141 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4201 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4261 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4321 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     4381 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4441 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4501 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4561 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4621 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4681 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     4741 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4801 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4861 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4921 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4981 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5041 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5101 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     5161 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5221 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     5281 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5341 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5401 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5461 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5521 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5581 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5641 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5701 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5761 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5821 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5881 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5941 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6001 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6061 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6121 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     6181 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6241 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6301 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6361 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6421 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6481 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6541 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6601 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6661 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6721 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6781 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6841 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6901 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6961 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7021 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7081 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7141 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7201 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7261 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7321 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7381 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7441 cacatttccc cgaaaagtgc cacctgacgt c
//
