LOCUS       sequence-469801-        6851 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of bovine arrestin-2 (R169E).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6851)
  AUTHORS   Pan L, Gurevich EV, Gurevich VV
  TITLE     The nature of the arrestin x receptor complex determines the 
            ultimate fate of the internalized receptor.
  JOURNAL   J Biol Chem. 2003 Mar 28;278(13):11623-32. doi: 
            10.1074/jbc.M209532200. Epub 2003 Jan 13.
  PUBMED    12525498
REFERENCE   2  (bases 1 to 6851)
  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.M209532200"; journalName: "J Biol Chem"; date: 
            "2003-03-28- 28"; volume: "278"; issue: "13"; pages: "11623-32"
COMMENT     Sequence Label: p3A2-RE
FEATURES             Location/Qualifiers
     source          1..6851
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     16..35
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(95..111)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(95..111)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(108..130)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    119..135
                     /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(143..149,150..167,168..173))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    188..209
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(326..343)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(497..1085)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(577..596)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1256..2047,2048..2116))
                     /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     1879..1898
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2117..2221)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2296..2315)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2487..2866
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2867..3070
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3021..3041
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3115..3134
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3115..3133
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        3225..3243
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     promoter        complement(4656..4674)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4694..4711)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4700..4924
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4970..5398
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5057..5076)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5267..5288
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5407..5427)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5412..5741
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5592..5727
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5654..5673
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5808..6602
                     /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(5862..5881)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6472..6491
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    join(6776..6851,1..46)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6813..6832)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
       61 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
      121 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
      181 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
      241 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
      301 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
      361 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
      421 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
      481 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
      541 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
      601 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
      661 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
      721 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
      781 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
      841 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
      901 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
      961 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1021 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     1081 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     1141 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     1201 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     1261 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     1321 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     1381 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     1441 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     1501 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     1561 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     1621 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     1681 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     1741 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     1801 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     1861 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     1921 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     1981 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     2041 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     2101 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     2161 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     2221 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     2281 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     2341 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     2401 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
     2461 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
     2521 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     2581 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     2641 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     2701 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     2761 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     2821 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     2881 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     2941 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     3001 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     3061 aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac
     3121 gactcactat agggagaccc aagcttggta ccgagctcgg atccactagt aacggccgcc
     3181 agtgtgctgg aattctgcag atatccatca cactggcggc cgcgatttag gtgacactat
     3241 agaatacaag aattcttttt tttttaaacc accatgggcg acaaagggac gcgggtgttc
     3301 aagaaggcga gccccaatgg aaagctcacc gtctatctgg gaaagcggga ctttgtggac
     3361 cacatcgacc tcgtggagcc cgtggatgga gtggttcttg tggatccgga gtatctcaag
     3421 gagaggagag tctatgtgac gctgacctgc gccttccgct acggccggga ggacctggat
     3481 gtcctgggcc tgacctttcg caaggacctg tttgtggcca acgtgcagtc tttcccgccg
     3541 gcccctgagg acaagaagcc cctgacgcgg ctgcaggagc gcctcatcaa gaagctgggc
     3601 gagcatgcct accctttcac ctttgagatc cctccgaacc tcccatgctc tgtgactttg
     3661 cagccgggac ctgaagatac agggaaggcc tgcggtgtgg actacgaagt gaaagccttc
     3721 tgtgcggaga acctggagga gaagatccac aagcggaatt ctgtgcgcct ggtcattgag
     3781 aaggttcagt atgccccaga gaggcctggc ccccagccca cggccgagac caccaggcag
     3841 ttcctcatgt cagacaagcc cttgcatctg gaggcctccc tggacaagga gatctactac
     3901 cacggagaac ccatcagtgt caacgtccat gtcaccaaca acaccaacaa gacggtgaag
     3961 aagatcaaga tctcggtgcg ccagtatgca gacatctgtc tgttcaacac agcccagtac
     4021 aagtgccctg tggccatgga agaggctgat gacacagtgg cacccagctc tacgttctgc
     4081 aaggtctaca cgctgacccc cttcctggcc aacaatcgag agaagcgggg cctcgccctg
     4141 gacgggaagc tcaaacacga ggacacgaac ctggcctcca gcaccctgtt gagggaagga
     4201 gccaaccggg agatcctggg catcattgtt tcctacaaag tgaaagtgaa gctggtggtg
     4261 tctcgtggcg gcctgttggg agatcttgca tccagtgatg tggccgtgga actgcctttc
     4321 accctaatgc accccaagcc caaagaggaa cccccacacc gggaagttcc agagcacgag
     4381 acgccggtag ataccaatct catagaactt gacaccaacg atgacgacat tgtgtttgag
     4441 gactttgccc gccagagact aaaaggcatg aaggatgaca aggaggaaga ggaggatggt
     4501 accggctctc cgcggctcaa cgacagatag actggggctg ccctccctcc gggcagctcc
     4561 aggtccactc tcatgcacta ggatgcttgt tcgtcttctt cctgtcctgg ctccccctcc
     4621 cctttgttct tccagtttct accagggggc cctattctat agtgtcacct aaatgctaga
     4681 gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     4741 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     4801 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     4861 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     4921 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     4981 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     5041 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     5101 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     5161 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     5221 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     5281 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     5341 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     5401 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     5461 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     5521 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     5581 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     5641 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     5701 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     5761 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     5821 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     5881 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     5941 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     6001 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     6061 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     6121 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     6181 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     6241 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     6301 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     6361 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     6421 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     6481 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     6541 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     6601 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     6661 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     6721 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     6781 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     6841 agcatttttt t
//
