LOCUS       sequence-469820-        7348 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of bovine arrestin-3 (deleted residues 176, 
            177, 178, 182-185).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7348)
  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 7348)
  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-delta7
FEATURES             Location/Qualifiers
     source          1..7348
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     246..265
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(484..588)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(663..682)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        854..1233
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1234..1437
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1388..1408
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1482..1501
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1482..1500
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(1535..1537,1538..1540,1541..2251)
                     /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(1580..1601)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1841..1860)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2188..2209
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(3520..3538)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3521..3538)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3558..3575)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3564..3788
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3834..4262
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3921..3940)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4131..4152
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4271..4291)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4276..4605
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4456..4591
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4518..4537
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4672..5466
                     /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(4726..4745)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5336..5355
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5640..5761
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5677..5696)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5731..5750
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5810..5826)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5810..5826)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5823..5845)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5834..5850
                     /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(5858..5864,5865..5882,5883..5888))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5903..5924
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6041..6058)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6212..6800)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6292..6311)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6971..7348,1..414,415..483))
                     /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"
ORIGIN
        1 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
       61 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
      121 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
      181 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
      241 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
      301 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
      361 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
      421 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
      481 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
      541 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
      601 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc
      661 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
      721 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
      781 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
      841 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
      901 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      961 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     1021 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     1081 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     1141 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     1201 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
     1261 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     1321 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     1381 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc
     1441 tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg
     1501 gagacccaag ctggaattcg cgatcgcggc cacgatggtg agcaagggcg aggagctgtt
     1561 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     1621 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctgatctg
     1681 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcct
     1741 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     1801 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     1861 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1921 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     1981 caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg
     2041 ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat
     2101 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccaaactgag
     2161 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     2221 gatcactctc ggcatggacg agctgtacaa gtccggactt aagtctcggc gagcgctcga
     2281 ctccatgggg gagaaacccg ggaccagggt cttcaagaaa tcgagtccta actgcaagct
     2341 caccgtgtac ctgggcaagc gggactttgt ggaccacctg gacaaagtgg atcctgtaga
     2401 tggtgtggtg ctagtggacc cggactactt gaaggaccgc aaagtgtttg tgaccctcac
     2461 ctgcgccttt cgctatggcc gcgaagacct ggacgtgctg ggcttgtcct tccgcaaaga
     2521 cctgttcatc gccaactacc aggccttccc cccaacaccc aacccacccc ggccccccac
     2581 ccgcctgcag gagcggctgc tgaggaagct gggccagcat gcccacccct tttttttcac
     2641 aataccccag aacctgccct gctccgtcac cctgcagcca ggcccagagg atacggggaa
     2701 ggcctgcggg gtcgactttg agattcgagc cttctgcgcc aaatcactag aagagaaaag
     2761 ccacaaacgg aactctgtgc gtctggtgat ccgaaaggtg cagtttccac ccggccccga
     2821 aaccacacgc cacttcctca tgtctgaccg gtccctgcac cttgaggctt cactggacaa
     2881 ggagctgtac taccacgggg agcccctcaa tgtcaatgtc cacgtcacca ataactccac
     2941 caagactgtc aagaagatca aagtctctgt gagacagtac gccgacatct gcctcttcag
     3001 caccgcccag tacaagtgcc ccgtggctca ggtcgaacaa gatgaccagg tgtcgcccag
     3061 ttccacgttc tgtaaggtgt acaccataac cccgctgctc agcaacaacc gggagaagcg
     3121 cggcctcgct ctagatggga agctcaagca cgaggacacc aacctggcct cgagcaccat
     3181 tgtgaaggag ggtgccaaca aggaggtgct ggggatactg gtgtcctaca gggtcaaggt
     3241 gaagctggtg gtgtcacgag gcggggatgt ctcagtggag ctgccatttg ttctcatgca
     3301 ccccaagccc catgatcaca tcgcccttcc caggccccag tcagctgttc ctgaaacaga
     3361 tgcccccgtg gacaccaacc tcattgaatt tgaaaccaac tatgccacag acgacgacat
     3421 cgtgttcgaa gactttgccc ggctgcggct caaggggctg aaggacgagg actatgacga
     3481 ccagttctgt atcgattagg gctggaagct tggccctatt ctatagtgtc acctaaatgc
     3541 tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3601 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3661 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3721 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     3781 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     3841 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3901 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3961 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4021 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4081 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4141 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4201 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4261 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4321 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4381 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4441 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4501 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4561 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4621 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     4681 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4741 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4801 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     4861 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4921 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4981 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     5041 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     5101 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5161 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     5221 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5281 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5341 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5401 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5461 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5521 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5581 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     5641 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5701 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5761 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     5821 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5881 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5941 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     6001 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     6061 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     6121 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     6181 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6241 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6301 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6361 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6421 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6481 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6541 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6601 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6661 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6721 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6781 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6841 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6901 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6961 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7021 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7081 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7141 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7201 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7261 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7321 tggcttcatt cagctccggt tcccaacg
//
