LOCUS       Exported                7530 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  plasma membrane marker.
ACCESSION   .
VERSION     .
KEYWORDS    TagBFP2-CAAX
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7530)
  AUTHORS   Goulden BD, Pacheco J, Dull A, Zewe JP, Deiters A, Hammond GRV
  TITLE     A high-avidity biosensor reveals plasma membrane PI(3,4)P2 is 
            predominantly a class I PI3K signaling product.
  JOURNAL   J Cell Biol. 2019 Mar 4;218(3):1066-1079. doi: 
            10.1083/jcb.201809026. Epub 2018 Dec 27.
  PUBMED    30591513
REFERENCE   2  (bases 1 to 7530)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7530
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             613..1314
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     polyA_signal    4318..4439
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4355..4374)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4409..4428
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4446..4901)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4583..4604)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4795..4814
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4928..5032
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5034..5391
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(5057..5077)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5242..5377
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5304..5323
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5426..6220
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5480..5499)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6090..6109
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6408..6427)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6452..6499
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6828..7416
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7317..7336
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag cgagctgatt aaggagaaca tgcacatgaa gctgtacatg
      661 gagggcaccg tggacaacca tcacttcaag tgcacatccg agggcgaagg caagccctac
      721 gagggcaccc agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc
      781 gacatcctgg ctactagctt cctctacggc agcaagacct tcatcaacca cacccagggc
      841 atccccgact tcttcaagca gtccttccct gagggcttca catgggagag agtcaccaca
      901 tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc
      961 atctacaacg tcaagatcag aggggtgaac ttcacatcca acggccctgt gatgcagaag
     1021 aaaacactcg gctgggaggc cttcaccgag acgctgtacc ccgctgacgg cggcctggaa
     1081 ggcagaaacg acatggccct gaagctcgtg ggcgggagcc atctgatcgc aaacgccaag
     1141 accacatata gatccaagaa acccgctaag aacctcaaga tgcctggcgt ctactatgtg
     1201 gactacagac tggaaagaat caaggaggcc aacaacgaga cctacgtcga gcagcacgag
     1261 gtggcagtgg ccagatactg cgacctccct agcaaactgg ggcacaagct taattccgga
     1321 ctcagatctc gagctcaagc ttcgaattct atggaaatta aagaggaagg ggcatcagaa
     1381 gaagggcagc actttcttcc tacagcccag gccaatgatc ccggggactg tcagttcaca
     1441 agtatccaga agactccaaa tgaaccgcag ttggaattca tccttgcatg caaggatctc
     1501 gtggctcctg tccgtgatcg taaactgaat acactggtgc agatctccgt aatccacccc
     1561 gtggagcaga gtctgacaag atactccagc accgaaattg tggagggaac aagggaccca
     1621 ctgtttttga ctggtgtcac attcccatct gagtatccca tctatgagga gaccaaaata
     1681 aaactaacag tctatgatgt caaggataag tctcatgaca ccgttcgaac cagtgtccta
     1741 ccagaacata aggatccccc gccagaagtt gggcgaagtt tcttgggcta tgccagtttt
     1801 aaagtgggag agctgctgaa gtcaaaggag caattgctgg tcctgagcct gagaacttca
     1861 gatggtggca aagtggttgg caccatagaa gtcagtgtcg tgaagatggg ggagattgag
     1921 gatggggaag ccgaccacat caccacagat gtacagggac aaaagtgtgc cctggtatgt
     1981 gaatgtacag ccccggaaag tgtgagcgga aaagataact tacctttttt gaattcagtg
     2041 ttaaagaacc cagtatgtaa attatataga tttcccacat ctgacaataa gtggatgcga
     2101 attcgagagc agatgtcaga gagcattctt tcctttcata ttcctaagga attgatttcc
     2161 cttcacatta aagaagattt gtgcagaaac caggagataa aagaacttgg tgagctttct
     2221 ccacattggg acaatctgcg aaaaaatgtc cttactcact gtgatcaaat ggtgaatatg
     2281 taccaagaca ttctgacaga acttagcaag gaaacagggt cctctttcaa atcaagcagc
     2341 agcaaaggag agaaaacatt agaatttgtt ccaataaatc tacatctgca aagaatgcag
     2401 gtacacagcc ctcacttgaa agatgctctc tacgatgtca tcactgtggg agccccagct
     2461 gcccattttc agggatttaa gaatggtggt cttcggaagc tactccatag atttgaaaca
     2521 gaaagaagaa ataccggata ccagtttatt tactattcac ctgaaaacac agccaaagca
     2581 aaggaagttc tcagcaacat caatcaacta caacctctta tagcaaccca tgcagaccta
     2641 ctgcttaatt ctgcaagcca gcattctcca gacagcttga agaattcttt aaagatgctt
     2701 tcagaaaaaa cagagctttt tgtacatgcc ttcaaggatc aacttgtcag gagtgctctt
     2761 ttagcactct acactgcaag gccaggaggc attcttaaga agccaccctc tcctaagagc
     2821 agcacagagg agagcagtcc ccaagaccaa cccccagtga tgagagggca ggactccata
     2881 ccacatcatt cagactatga tgaggaagag tgggacaggg tgtgggccaa tgtggggaag
     2941 agcctgaact gcattattgc tatggtggac aaactgattg aaagagatgg tggcagtgaa
     3001 ggcagtggcg gcaacaatga tggagaaaag gaaccttcat taacagatgc cattccctct
     3061 cacccaagag aggactggta tgaacagttg tatcccctca tccttaccct gaaggactgc
     3121 atgggagaag tggtgaaccg agccaagcag tccctgacat ttgtgctcct tcaggaactt
     3181 gcgtacagct tgccccagtg tctgatgctg acgctaagaa gagacatcgt cttcagccaa
     3241 gcacttgctg gattggtttg tggttttatc atcaaattac agacaagtct gtatgaccca
     3301 ggcttcctac agcagcttca cacagtgggg ttgatagtac aatatgaagg actgctaagt
     3361 acatacagcg atgaaattgg aatgctagag gacatggccg ttggcatttc cgatttaaag
     3421 aaagttgcat ttaaaataat tgaagccaaa tccaatgatg tgttgccagt tataacagga
     3481 agacgagaac attacgtggt agaggtcaag cttccagcca gaatgtttga gtcactacct
     3541 ctacagatta aagaaggaca gttgcttcat gtgtatccag tactttttaa tgttggaatc
     3601 aatgaacagc aaactctggc tgaaaggttt ggagatgtct ctttgcaaga aagtattaat
     3661 caggaaaact tcgaacttct acaagaatat tacaagatat ttatggaaaa gatgcctcct
     3721 gattatattt cacattttca ggaacaaaat gatttaaaag cattgctaga aaatctcctt
     3781 caaaatatcc aatccaaaaa aagaaagaat gtagaaatta tgtggctggc tgcaacgatt
     3841 tgccgcaaac tgaatggtat tcgtttcacc tgttgtaaaa gtgccaaaga caggacatcg
     3901 atgtcagtga cacttgaaca atgctcaatc ttgagagatg agcaccagtt acacaaggac
     3961 ttctttatcc gagcgctgga ttgcatgaga agagaaggat gccgcataga gaatgtactg
     4021 aagaatatca aatgcagaaa gtatgctttc aacatgctac agctgatggc tttccccaag
     4081 tactacagac ctccagaggg gacttatgga aaagctgaca ccgcagtcga caaccctcct
     4141 gatgagagtg gccccggctg catgagctgc aagtgtgtgc tctcctgagg atccaccgga
     4201 tctagataac tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     4261 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     4321 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     4381 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     4441 cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     4501 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     4561 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     4621 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     4681 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     4741 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     4801 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     4861 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     4921 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4981 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     5041 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     5101 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     5161 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     5221 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     5281 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     5341 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     5401 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     5461 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     5521 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     5581 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     5641 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     5701 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     5761 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     5821 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     5881 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     5941 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     6001 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     6061 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     6121 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     6181 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     6241 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     6301 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     6361 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     6421 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     6481 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     6541 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     6601 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     6661 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6721 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6781 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6841 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6901 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6961 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     7021 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7081 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7141 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7201 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7261 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7321 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7381 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7441 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     7501 tctgtggata accgtattac cgccatgcat
//
