LOCUS       sequence-224756-        4758 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  plasma membrane marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4758)
  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 4758)
  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: 
            "10.1083/jcb.201809026"; journalName: "J Cell Biol"; date: 
            "2019-03-4- 4"; volume: "218"; issue: "3"; pages: "1066-1079"
COMMENT     Sequence Label: TagBFP2-CAAX
FEATURES             Location/Qualifiers
     source          1..4758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        37..394
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        37..228
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(60..80)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      245..380
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             429..1223
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             429..1223
                     /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(483..502)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1093..1112
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1411..1430)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1455..1502
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1831..2419
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2320..2339
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2594..2897
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2898..3101
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3052..3072
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3140..3149
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3146..3847
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     polyA_signal    4079..4200
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4116..4135)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4170..4189
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4207..4662)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4344..4365)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4556..4575
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(4689..4758,1..35)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
       61 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
      121 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
      181 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
      241 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
      301 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
      361 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
      421 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
      481 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
      541 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
      601 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
      661 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
      721 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
      781 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
      841 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
      901 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
      961 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     1021 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     1081 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     1141 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     1201 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     1261 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     1321 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     1381 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     1441 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     1501 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     1561 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     1621 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     1681 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     1741 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     1801 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     1861 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     1921 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     1981 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2041 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2101 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2161 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2221 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2281 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2341 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2401 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2461 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     2521 taccgccatg cattagttat taatagtaat caattacggg gtcattagtt catagcccat
     2581 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     2641 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     2701 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     2761 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     2821 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     2881 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     2941 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     3001 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     3061 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
     3121 tccgctagcg ctaccggtcg ccaccatggt gagcgagctg attaaggaga acatgcacat
     3181 gaagctgtac atggagggca ccgtggacaa ccatcacttc aagtgcacat ccgagggcga
     3241 aggcaagccc tacgagggca cccagaccat gagaatcaag gtggtcgagg gcggccctct
     3301 ccccttcgcc ttcgacatcc tggctactag cttcctctac ggcagcaaga ccttcatcaa
     3361 ccacacccag ggcatccccg acttcttcaa gcagtccttc cctgagggct tcacatggga
     3421 gagagtcacc acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcctcca
     3481 ggacggctgc ctcatctaca acgtcaagat cagaggggtg aacttcacat ccaacggccc
     3541 tgtgatgcag aagaaaacac tcggctggga ggccttcacc gagacgctgt accccgctga
     3601 cggcggcctg gaaggcagaa acgacatggc cctgaagctc gtgggcggga gccatctgat
     3661 cgcaaacgcc aagaccacat atagatccaa gaaacccgct aagaacctca agatgcctgg
     3721 cgtctactat gtggactaca gactggaaag aatcaaggag gccaacaacg agacctacgt
     3781 cgagcagcac gaggtggcag tggccagata ctgcgacctc cctagcaaac tggggcacaa
     3841 gcttaattcc ggactcagat ctcgagctca agcttcgaat tctgcagtcg acaaccctcc
     3901 tgatgagagt ggccccggct gcatgagctg caagtgtgtg ctctcctgag gatccaccgg
     3961 atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     4021 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     4081 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4141 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4201 acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     4261 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     4321 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     4381 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     4441 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     4501 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     4561 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     4621 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     4681 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4741 tcatgagaca ataaccct
//
