LOCUS       sequence-313688-        6822 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ExRai-AKAR2 targeted to the cytosolic face of the ER membrane..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6822)
  AUTHORS   Zhang JF, Liu B, Hong I, Mo A, Roth RH, Tenner B, Lin W, Zhang JZ, 
            Molina RS, Drobizhev M, Hughes TE, Tian L, Huganir RL, Mehta S, 
            Zhang J
  TITLE     An ultrasensitive biosensor for high-resolution kinase activity 
            imaging in awake mice.
  JOURNAL   Nat Chem Biol. 2020 Sep 28. pii: 10.1038/s41589-020-00660-y. doi: 
            10.1038/s41589-020-00660-y.
  PUBMED    32989297
REFERENCE   2  (bases 1 to 6822)
  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: "Nat Chem 
            Biol. 2020 Sep 28. pii: 10.1038/s41589-020-00660-y. doi: 
            10.1038/s41589-020-00660-y."
COMMENT     Sequence Label: pcDNA3-erExRai-AKAR2
FEATURES             Location/Qualifiers
     source          1..6822
                     /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"
     primer_bind     1291..1312
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1418..1439)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1679..1698)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     promoter        complement(2303..2321)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2304..2321)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2341..2358)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2347..2571
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2617..3045
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2704..2723)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2914..2935
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3054..3074)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3144..3460
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3311..3446
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3373..3392
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3527..4321
                     /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(3581..3600)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4191..4210
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4495..4616
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4532..4551)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4586..4605
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4665..4681)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4665..4681)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4678..4700)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4689..4705
                     /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(4713..4719,4720..4737,4738..4743))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4758..4779
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4896..4913)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5067..5655)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5147..5166)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5826..6617,6618..6686))
                     /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     6449..6468
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6687..6791)
                     /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 gcttggtacc
      901 ccttgcagct tccagttgtc aggaagagaa ggctcccatg gaccctgtgg tggtgctggg
      961 gctctgtctc tcctgtttgc ttctcctttc actctggaaa cagagctatg ggggagggga
     1021 tcccatgctg cgtcgcgcca ccctggtgga cggcggcacc ggcggcagcg agctcagctt
     1081 ctgcaacgtc tatatcaagg ccgacaagca gaagaacggc atcaaggcga acttcaagat
     1141 ccgccacaac atcgaggacg gcggcgtgca gctcgcctac cactaccagc agaacacccc
     1201 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcgtgc agtccaaact
     1261 ttcgaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     1321 cgggatcact ctcggcatgg acgagctgta caagggcggt accggaggga gcatggtgag
     1381 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     1441 aaacggccac aagttcagcg tgtccggcga gggtgagggc gatgccacct acggcaagct
     1501 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     1561 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     1621 cttcttcaag tccgccatgc ccgaaggcta catccaggag cgcaccatct tcttcaagga
     1681 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1741 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1801 gtacaacttg ctgcataagt tttctcaaga acagatcggc gaaaacattg tgtgcagggt
     1861 catttgtacc acgggtcaaa ttcccatccg agatttgtca gctgatattt cacaagtgct
     1921 taaggaaaaa cgatccataa agaaagtttg gacatttggt agaaacccag cctgtgacta
     1981 tcatttagga aacatttcaa gactgtcaaa taagcatttc caaatactac taggagaaga
     2041 cggtaacctt ttattgaatg acatttccac taatgggacc tggttaaatg ggcaaaaagt
     2101 cgagaagaac agcaatcagt tactgtctca aggtgatgaa ataaccgttg gtgtaggcgt
     2161 ggaatcagat attttatctc tggtcatttt cataaacgac aaatttaagc agtgcctcga
     2221 gcagaacaaa gttgatcgct aggaattctg cagatatcca tcacactggc ggccgctcga
     2281 gcatgcatct agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag
     2341 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     2401 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     2461 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     2521 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     2581 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     2641 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     2701 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     2761 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     2821 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     2881 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     2941 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     3001 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     3061 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3121 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3181 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3241 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3301 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3361 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     3421 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     3481 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3541 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3601 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3661 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     3721 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     3781 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3841 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     3901 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3961 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4021 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     4081 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4141 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4201 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4261 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4321 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     4381 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     4441 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     4501 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4561 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     4621 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     4681 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     4741 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     4801 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     4861 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4921 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4981 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5041 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5101 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5161 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5221 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5281 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5341 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5401 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5461 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5521 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5581 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5641 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5701 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5761 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5821 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5881 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5941 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6001 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6061 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6121 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6181 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6241 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6301 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6361 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6421 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6481 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6541 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6601 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6661 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6721 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6781 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
