LOCUS       sequence-469798-        7369 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of bovine arrestin-3 (R170E).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7369)
  AUTHORS   Zheng C, Weinstein LD, Nguyen KK, Grewal A, Gurevich EV, Gurevich VV
  TITLE     GPCR Binding and JNK3 Activation by Arrestin-3 Have Different 
            Structural Requirements.
  JOURNAL   Cells. 2023 Jun 6;12(12):1563. doi: 10.3390/cells12121563.
  PUBMED    37371033
REFERENCE   2  (bases 1 to 7369)
  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"; journalName: "Cells."; 
            date: "2023-06-6"; volume: "12(12):1563. doi"; pages: " 
            10.3390/cells12121563"
COMMENT     Sequence Label: pVEA3-RE
FEATURES             Location/Qualifiers
     source          1..7369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      433..1021
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     922..941
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1175..1192
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1309..1330
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1345..1350,1351..1368,1369..1375)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1383..1399
                     /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     1388..1410
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1407..1423
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1407..1423
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1472..1593
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1483..1502)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1537..1556
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1767..2561)
                     /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(1878..1897)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2488..2507
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2628..2957)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2642..2777
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2696..2715)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2942..2962
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2971..3399)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3081..3102)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3293..3312
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3445..3669
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3658..3675
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        3695..3713
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3695..3712
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(join(5003..5713,5714..5716,5717..5719))
                     /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(5045..5066)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5394..5413
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5653..5674
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5753..5772)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5754..5772)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(5817..6020)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5846..5866)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6021..6400
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     6572..6591
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6666..6770
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6771..6839,6840..7369,1..262)
                     /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(6989..7008)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
       61 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
      121 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
      181 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
      241 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
      301 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
      361 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
      421 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
      481 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
      541 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
      601 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
      661 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
      721 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
      781 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
      841 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
      901 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
      961 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     1021 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     1081 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     1141 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     1201 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     1261 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     1321 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     1381 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct
     1441 ctagctagag gtcgacggta tacagacatg ataagataca ttgatgagtt tggacaaacc
     1501 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     1561 tttgtaacca ttataagctg caataaacaa gttggggtgg gcgaagaact ccagcatgag
     1621 atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct
     1681 ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc
     1741 ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg
     1801 atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg
     1861 ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc
     1921 acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc
     1981 ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg
     2041 agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga
     2101 tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg
     2161 tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg
     2221 gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc
     2281 aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg
     2341 cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg
     2401 gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg
     2461 gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa
     2521 gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct
     2581 gtctcttgat cagatccgaa aatggatata caagctcccg ggagcttttt gcaaaagcct
     2641 aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggcaga ggcggcctcg
     2701 gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc
     2761 ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga
     2821 tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca cctggttgct
     2881 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
     2941 caccctaact gacacacatt ccacagaatt aattcgcgtt aaatttttgt taaatcagct
     3001 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     3061 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     3121 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     3181 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     3241 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     3301 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     3361 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtg gggatacccc ctagagcccc
     3421 agctggttct ttccgcctca gaagccatag agcccaccgc atccccagca tgcctgctat
     3481 tgtcttccca atcctccccc ttgctgtcct gccccacccc accccccaga atagaatgac
     3541 acctactcag acaatgcgat gcaatttcct cattttatta ggaaaggaca gtgggagtgg
     3601 caccttccag ggtcaaggaa ggcacggggg aggggcaaac aacagatggc tggcaactag
     3661 aaggcacagt cgaggctgat cagcgagctc tagcatttag gtgacactat agaatagggc
     3721 caagcttcca gccctaatcg atacagaact ggtcgtcata gtcctcgtcc ttcagcccct
     3781 tgagccgcag ccgggcaaag tcttcgaaca cgatgtcgtc gtctgtggca tagttggttt
     3841 caaattcaat gaggttggtg tccacggggg catctgtttc aggaacagct gactggggcc
     3901 tgggaagggc gatgtgatca tggggcttgg ggtgcatgag aacaaatggc agctccactg
     3961 agacatcccc gcctcgtgac accaccagct tcaccttgac cctgtaggac accagtatcc
     4021 ccagcacctc cttgttggca ccctccttca caatggtgct cgaggccagg ttggtgtcct
     4081 cgtgcttgag cttcccatct agagcgaggc cgcgcttctc ccggttgttg ctgagcagcg
     4141 gggttatggt gtacacctta cagaacgtgg aactgggcga cacctggtca tcttgttcga
     4201 cctgagccac ggggcacttg tactgggcgg tgctgaagag gcagatgtcg gcgtactgtc
     4261 tcacagagac tttgatcttc ttgacagtct tggtggagtt attggtgacg tggacattga
     4321 cattgagggg ctccccgtgg tagtacagct ccttgtccag tgaagcctca aggtgcaggg
     4381 accggtcaga catgaggaag tggcgtgtgg tttcagctga aggctggggc cctggtttct
     4441 ctggggcaaa ctgcaccttt tcgatcacca gacgtacgga gttccgtttg tggcttttct
     4501 cttctagtga tttggcgcag aaggctcgaa tctcaaagtc gaccccgcaa gccttccccg
     4561 tatcctctgg gcctggctgt agggtgacgg agcagggcag gttctggggt attgtgaaaa
     4621 aaaaggggtg ggcatgctgg cccagcttcc tcagcagccg ctcctgcaga cgcgtaggtg
     4681 gtctaggtgg gttaggtgtt ggagggaagg cctggtagtt ggcgatgaac aggtctttgc
     4741 ggaaggacaa gcccaggacg tccaggtctt cgcggccata gcgaaaggcg cacgtgaggg
     4801 tcacaaacac tttgcggtcc ttcaagtagt ccggatccac tagcaccaca ccatctacag
     4861 ggtccacttt gtccaggtgg tccacaaagt cccgcttgcc taggtacacg gtgagcttgc
     4921 agttaggact cgatttcttg aagaccctgg tcccgggttt ctcccccatg gagtcgagcg
     4981 ctcgccgaga cttaagtccg gacttgtaca gctcgtccat gccgagagtg atcccggcgg
     5041 cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcagtt
     5101 tggactggta gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg
     5161 tgttctgctg gtagtggtcg gcgagctgca cgccgccgtc ctcgatgttg tggcggatct
     5221 tgaagttggc cttgatgccg ttcttctgct tgtcggcggt gatatagacg ttgtggctgt
     5281 tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct
     5341 tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt
     5401 agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg
     5461 acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg ggcgaagcac tgcaggccgt
     5521 agcccagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatca
     5581 gcttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact
     5641 tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct
     5701 cctcgccctt gctcaccatc gtggccgcga tcgcgaattc cagcttgggt ctccctatag
     5761 tgagtcgtat taatttcgat aagccagtaa gcagtgggtt ctctagttag ccagagagct
     5821 ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag
     5881 ttgttacgac attttggaaa gtcccgttga ttttggtgcc aaaacaaact cccattgacg
     5941 tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg cccattgatg
     6001 tactgccaaa accgcatcac catggtaata gcgatgacta atacgtagat gtactgccaa
     6061 gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca tttaccgtca
     6121 ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc caagtgggca
     6181 gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg cgttactatg
     6241 ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg
     6301 ccatttaccg taagttatgt aacgcggaac tccatatatg ggctatgaac taatgacccc
     6361 gtaattgatt actattaata actagtcaat aatcaatgtc aacgcgtata tctggcccgt
     6421 acatcgcgaa gcagcgcaaa acgcctaacc ctaagcagat tcttcatgca attgtcggtc
     6481 aagccttgcc ttgttgtagc ttaaattttg ctcgcgcact actcagcgac ctccaacaca
     6541 caagcaggga gcagatactg gcttaactat gcggcatcag agcagattgt actgagagtg
     6601 caccataggg gatcgggaga tctcccgatc cgtcgacgtc aggtggcact tttcggggaa
     6661 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     6721 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     6781 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     6841 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     6901 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     6961 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     7021 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     7081 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     7141 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     7201 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     7261 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     7321 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagc
//
