LOCUS       sequence-473894-        7450 bp    DNA     circular SYN 27-MAY-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 7450)
  AUTHORS   Hanson SM, Cleghorn WM, Francis DJ, Vishnivetskiy SA, Raman D, Song 
            X, Nair KS, Slepak VZ, Klug CS, Gurevich VV
  TITLE     Arrestin mobilizes signaling proteins to the cytoskeleton and 
            redirects their activity.
  JOURNAL   J Mol Biol. 2007 Apr 27;368(2):375-87. doi: 
            10.1016/j.jmb.2007.02.053. Epub 2007 Feb 22.
  PUBMED    17359998
REFERENCE   2  (bases 1 to 7450)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 27, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.jmb.2007.02.053"; journalName: "J Mol Biol"; date: 
            "2007-04-27- 27"; volume: "368"; issue: "2"; pages: "375-87"
COMMENT     Sequence Label: pVeBtA2d7
FEATURES             Location/Qualifiers
     source          1..7450
                     /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(3003..3021)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3004..3021)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3041..3058)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3047..3271
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3317..3745
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3404..3423)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3614..3635
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3754..3774)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3759..4088
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3939..4074
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4001..4020
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4155..4949
                     /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(4209..4228)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4819..4838
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5123..5244
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5160..5179)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5214..5233
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5293..5309)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5293..5309)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5306..5328)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5317..5333
                     /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(5341..5347,5348..5365,5366..5371))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5386..5407
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5524..5541)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5695..6283)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5775..5794)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6454..7245,7246..7314))
                     /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     7077..7096
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7315..7419)
                     /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 gtatccaccc gggcccgaga ccaccaggca gttcctcatg
     2221 tcagacaagc ccttgcatct ggaggcctcc ctggacaagg agatctacta ccacggagaa
     2281 cccatcagtg tcaacgtcca tgtcaccaac aacaccaaca agacggtgaa gaagatcaag
     2341 atctcggtgc gccagtatgc agacatctgt ctgttcaaca cagcccagta caagtgccct
     2401 gtggccatgg aagaggctga tgacacagtg gcacccagct ctacgttctg caaggtctac
     2461 acgctgaccc ccttcctggc caacaatcga gagaagcggg gcctcgctct agacgggaag
     2521 ctcaaacacg aggacacgaa cctggcctcg agcaccctgt tgagggaagg agccaaccgg
     2581 gaaatcctgg gcatcattgt ttcctacaaa gtgaaagtga agctggtggt gtctcgtggc
     2641 ggcctgttgg gagatcttgc atccagtgat gtggccgtgg aactgccttt caccctaatg
     2701 caccccaagc ccaaagagga acccccacac cgggaagttc cagagcacga gacgccggta
     2761 gataccaatc tcatagaact tgacaccaac gatgacgaca ttgtgtttga ggactttgcc
     2821 cgcctgagac taaaaggcat gaaggatgac aaggaggaag aggaggatgg taccggctct
     2881 ccgcggctca acgacagaat cgattagcgg ccgctgccct ccctccgggc agctccaggt
     2941 ccactctcat gcactaggat gcttgttcgt cttcttcctg tcctggctaa gcttggccct
     3001 attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt gccttctagt
     3061 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3121 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     3181 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3241 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     3301 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3361 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3421 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3481 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3541 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3601 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3661 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     3721 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     3781 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3841 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3901 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3961 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4021 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4081 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     4141 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4201 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4261 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4321 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4381 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4441 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4501 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4561 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4621 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4681 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4741 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4801 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4861 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4921 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     4981 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5041 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5101 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5161 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5221 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5281 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5341 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5401 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5461 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5521 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5581 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5641 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5701 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5761 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5821 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5881 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5941 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6001 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6061 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6121 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6181 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6241 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6301 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6361 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6421 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6481 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6541 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6601 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6661 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6721 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6781 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6841 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6901 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6961 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7021 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7081 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7141 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7201 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7261 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7321 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7381 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7441 acctgacgtc
//
