LOCUS       sequence-469803-        7369 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of bovine arrestin-3.
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: p3VEA3
FEATURES             Location/Qualifiers
     source          1..7369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      626..1214
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1115..1134
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1368..1385
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1502..1523
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1538..1543,1544..1561,1562..1568)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1576..1592
                     /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     1581..1603
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1600..1616
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1600..1616
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1665..1786
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1676..1695)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1730..1749
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1960..2754)
                     /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(2071..2090)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2681..2700
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2821..3150)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2835..2970
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2889..2908)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     3135..3155
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(3164..3592)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3274..3295)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3486..3505
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3638..3862
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3851..3868
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        3888..3906
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3888..3905
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(join(5196..5906,5907..5909,5910..5912))
                     /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(5238..5259)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5587..5606
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5846..5867
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5946..5965)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5947..5965)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(6010..6213)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6039..6059)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6214..6593
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     6765..6784
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6859..6963
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6964..7032,7033..7369,1..455)
                     /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(7182..7201)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
       61 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
      121 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
      181 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
      241 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
      301 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
      361 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
      421 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
      481 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
      541 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
      601 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
      661 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
      721 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
      781 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
      841 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
      901 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
      961 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     1021 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     1081 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     1141 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     1201 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     1261 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     1321 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     1381 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     1441 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     1501 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     1561 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     1621 attacgccaa gctctagcta gaggtcgacg gtatacagac atgataagat acattgatga
     1681 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     1741 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttgggg tgggcgaaga
     1801 actccagcat gagatccccg cgctggagga tcatccagcc ggcgtcccgg aaaacgattc
     1861 cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa atctcgtgat ggcaggttgg
     1921 gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct cagaagaact cgtcaagaag
     1981 gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg
     2041 gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg
     2101 atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc
     2161 caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg
     2221 catgcgcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc
     2281 cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg
     2341 tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc
     2401 atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc
     2461 cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc
     2521 tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cctgcagttc
     2581 attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag
     2641 ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag
     2701 cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa
     2761 cgatcctcat cctgtctctt gatcagatcc gaaaatggat atacaagctc ccgggagctt
     2821 tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc
     2881 agaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg
     2941 ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc
     3001 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     3061 acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     3121 tggggacttt ccacacccta actgacacac attccacaga attaattcgc gttaaatttt
     3181 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     3241 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     3301 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     3361 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     3421 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     3481 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     3541 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtggggatac
     3601 cccctagagc cccagctggt tctttccgcc tcagaagcca tagagcccac cgcatcccca
     3661 gcatgcctgc tattgtcttc ccaatcctcc cccttgctgt cctgccccac cccacccccc
     3721 agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta ttaggaaagg
     3781 acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca aacaacagat
     3841 ggctggcaac tagaaggcac agtcgaggct gatcagcgag ctctagcatt taggtgacac
     3901 tatagaatag ggccaagctt ccagccctaa tcgatacaga actggtcgtc atagtcctcg
     3961 tccttcagcc ccttgagccg cagccgggca aagtcttcga acacgatgtc gtcgtctgtg
     4021 gcatagttgg tttcaaattc aatgaggttg gtgtccacgg gggcatctgt ttcaggaaca
     4081 gctgactggg gcctgggaag ggcgatgtga tcatggggct tggggtgcat gagaacaaat
     4141 ggcagctcca ctgagacatc cccgcctcgt gacaccacca gcttcacctt gaccctgtag
     4201 gacaccagta tccccagcac ctccttgttg gcaccctcct tcacaatggt gctcgaggcc
     4261 aggttggtgt cctcgtgctt gagcttccca tctagagcga ggccgcgctt ctcccggttg
     4321 ttgctgagca gcggggttat ggtgtacacc ttacagaacg tggaactggg cgacacctgg
     4381 tcatcttgtt cgacctgagc cacggggcac ttgtactggg cggtgctgaa gaggcagatg
     4441 tcggcgtact gtctcacaga gactttgatc ttcttgacag tcttggtgga gttattggtg
     4501 acgtggacat tgacattgag gggctccccg tggtagtaca gctccttgtc cagtgaagcc
     4561 tcaaggtgca gggaccggtc agacatgagg aagtggcgtg tggtttcagc tgaaggctgg
     4621 ggccctggtt tctctggggc aaactgcacc tttcggatca ccagacgtac ggagttccgt
     4681 ttgtggcttt tctcttctag tgatttggcg cagaaggctc gaatctcaaa gtcgaccccg
     4741 caagccttcc ccgtatcctc tgggcctggc tgtagggtga cggagcaggg caggttctgg
     4801 ggtattgtga aaaaaaaggg gtgggcatgc tggcccagct tcctcagcag ccgctcctgc
     4861 agacgcgtag gtggtctagg tgggttaggt gttggaggga aggcctggta gttggcgatg
     4921 aacaggtctt tgcggaagga caagcccagg acgtccaggt cttcgcggcc atagcgaaag
     4981 gcgcacgtga gggtcacaaa cactttgcgg tccttcaagt agtccggatc cactagcacc
     5041 acaccatcta cagggtccac tttgtccagg tggtccacaa agtcccgctt gcctaggtac
     5101 acggtgagct tgcagttagg actcgatttc ttgaagaccc tggtcccggg tttctccccc
     5161 atggagtcga gcgctcgccg agacttaagt ccggacttgt acagctcgtc catgccgaga
     5221 gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg
     5281 tctttgctca gtttggactg gtagctcagg tagtggttgt cgggcagcag cacggggccg
     5341 tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgccgcc gtcctcgatg
     5401 ttgtggcgga tcttgaagtt ggccttgatg ccgttcttct gcttgtcggc ggtgatatag
     5461 acgttgtggc tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg
     5521 aagtcgatgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg
     5581 gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct
     5641 tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcgggcgaag
     5701 cactgcaggc cgtagcccag ggtggtcacg agggtgggcc agggcacggg cagcttgccg
     5761 gtggtgcaga tcagcttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg
     5821 gacacgctga acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc
     5881 ccggtgaaca gctcctcgcc cttgctcacc atcgtggccg cgatcgcgaa ttccagcttg
     5941 ggtctcccta tagtgagtcg tattaatttc gataagccag taagcagtgg gttctctagt
     6001 tagccagaga gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt
     6061 caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa
     6121 actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc
     6181 acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta
     6241 gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg
     6301 ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac
     6361 tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat
     6421 tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg
     6481 tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg
     6541 aactaatgac cccgtaattg attactatta ataactagtc aataatcaat gtcaacgcgt
     6601 atatctggcc cgtacatcgc gaagcagcgc aaaacgccta accctaagca gattcttcat
     6661 gcaattgtcg gtcaagcctt gccttgttgt agcttaaatt ttgctcgcgc actactcagc
     6721 gacctccaac acacaagcag ggagcagata ctggcttaac tatgcggcat cagagcagat
     6781 tgtactgaga gtgcaccata ggggatcggg agatctcccg atccgtcgac gtcaggtggc
     6841 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     6901 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     6961 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     7021 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     7081 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     7141 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     7201 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     7261 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     7321 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttac
//
