LOCUS       sequence-243582-        7713 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SB-transposon plasmid for stable expression of lipid raft-targeted 
            Lyn-AktAR2 variant of FRET-based AKT activity reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7713)
  AUTHORS   Havranek O, Xu J, Kohrer S, Wang Z, Becker L, Comer JM, Henderson J,
            Ma W, Man Chun Ma J, Westin JR, Ghosh D, Shinners N, Sun L, Yi AF, 
            Karri AR, Burger JA, Zal T, Davis RE
  TITLE     Tonic B-cell receptor signaling in diffuse large B-cell lymphoma.
  JOURNAL   Blood. 2017 Aug 24;130(8):995-1006. doi: 
            10.1182/blood-2016-10-747303. Epub 2017 Jun 23.
  PUBMED    28646116
REFERENCE   2  (bases 1 to 7713)
  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: 
            "10.1182/blood-2016-10-747303"; journalName: "Blood"; date: 
            "2017-08-24- 24"; volume: "130"; issue: "8"; pages: "995-1006"
COMMENT     Sequence Label: pSBbi-Pur-Lyn-AktAR2
FEATURES             Location/Qualifiers
     source          1..7713
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1130..1150
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1208..1217
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1256..1258,1259..1261,1262..1972)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAIHGNVYITADKQKNGIK
                     ANFGLNCNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             2525..3259
                     /codon_start=1
                     /product="circularly permuted form of the Venus yellow 
                     fluorescent protein (Nagai et al., 2004)"
                     /label=cp173Venus
                     /note="efficient FRET acceptor for CFP"
                     /translation="MGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDH
                     MVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEELFTGVVPILVELDGDVNGHKFSVSG
                     EGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPE
                     GYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHN
                     VYITADKQKNGIKANFKIRHNIE"
     primer_bind     complement(3313..3330)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3319..3543
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        3975..4079
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4080..4148,4149..4940)
                     /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     complement(4298..4317)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5111..5699
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5600..5619
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5853..5870
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6244..6263
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    6262..6317
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     6363..6382
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(6482..7081)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6565..6585)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     7062..7081
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        join(7713,1..1182)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
ORIGIN
        1 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
       61 gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
      121 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
      181 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg
      241 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta
      301 cttccacgcc cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg
      361 tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc
      421 tggcttgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct
      481 gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc
      541 tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg
      601 gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg
      661 cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg
      721 cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca
      781 ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg
      841 aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt
      901 ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc
      961 gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg
     1021 atggagtttc cccatactga gtgggtggag actgaagtta ggccagcttg gcacttgatg
     1081 taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag
     1141 acagtggttc aaagtttttt tcttccattt aaggtgtcgt gaaaactacc ccaagctggc
     1201 ctctgaggcc accatggggt gcattaagtc taagcggaag gacaaaaagg atcccatggt
     1261 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1321 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     1381 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     1441 gaccaccctg agctggggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca
     1501 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1561 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1621 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1681 ggagtacaac gccatccacg gcaacgtcta tatcaccgcc gacaagcaga agaacggcat
     1741 caaggccaac ttcggcctca actgcaacat cgaggacggc agcgtgcagc tcgccgacca
     1801 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1861 gagcacccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     1921 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agcgcatgca
     1981 taagttttct caagaacaga tcggcgaaaa cattgtgtgc agggtcattt gtaccacggg
     2041 tcaaattccc atccgagatt tgtcagctga tatttcacaa gtgcttaagg aaaaacgatc
     2101 cataaagaaa gtttggacat ttggtagaaa cccagcctgt gactatcatt taggaaacat
     2161 ttcaagactg tcaaataagc atttccaaat actactagga gaagacggta accttttatt
     2221 gaatgacatt tccactaatg ggacctggtt aaatgggcaa aaagtcgaga agaacagcaa
     2281 tcagttactg tctcaaggtg atgaaataac cgttggtgta ggcgtggaat cagatatttt
     2341 atctctggtc attttcataa acgacaaatt taagcagtgc ctcgagcaga acaaagttga
     2401 tcgctctgca ggtaagccag gcagcggcga gggcagcacc aagggcctgg tcgaccctcg
     2461 tccgcgctcg tgcacctggc cggaccccag gccggagttt ggaggtaccg gcggcagcga
     2521 gctcatgggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     2581 ccccgtgctg ctgcccgaca accactacct gagctaccag tccaagctga gcaaagaccc
     2641 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     2701 cggcatggac gagctgtaca agggaggtac cggtggatct atggtgagca agggcgagga
     2761 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     2821 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct
     2881 gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta
     2941 cggccttcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     3001 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     3061 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     3121 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     3181 cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa
     3241 gatccgccac aacatcgagt aaggcctgtc aggccaagct tggggatcaa ttctctagag
     3301 ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     3361 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     3421 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     3481 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     3541 tggcttctga ggcggaaaga accagctggg cctactagat agcggacccc ttaccgaaac
     3601 atcgccgcat tctgcagagg agtcgaactt agtgtatgta aacttctgac ccactggaat
     3661 tgtgatacag tgaattataa gtgaaataat ctgtctgtaa acaattgttg gaaaaatgac
     3721 ttgtgtcatg cacaaagtag atgtcctaac tgacttgcca aaactattgt ttgttaacaa
     3781 gaaatttgtg gagtagttga aaaacgagtt ttaatgactc caacttaagt gtatgtaaac
     3841 ttccgacttc aactgtatag ggatccaatt ttaacaaaat attaacgctt acaatttcct
     3901 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatca ggtggcactt
     3961 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4021 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4081 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4141 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4201 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4261 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4321 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4381 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     4441 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     4501 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     4561 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     4621 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     4681 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     4741 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     4801 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     4861 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     4921 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     4981 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5041 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5101 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5161 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5221 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     5281 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5341 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5401 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5461 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5521 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5581 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5641 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5701 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     5761 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     5821 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggatc
     5881 cctatacagt tgaagtcgga agtttacata caccttagcc aaatacattt aaactcactt
     5941 tttcacaatt cctgacattt aatcctagta aaaattccct gtcttaggtc agttaggatc
     6001 accactttat tttaagaatg tgaaatatca gaataatagt agagagaatg attcatttca
     6061 gcttttattt ctttcatcac attcccagtg ggtcagaagt ttacatacac tcgactcctc
     6121 tgcagaatgc ggcgatgttt cggtaagggg tcccgagtag ctagttcatg gcagccagca
     6181 tatggcatat gttgccaaac tctaaaccaa atactcattc tgatgtttta aatgatttgc
     6241 cctcccatat gtccttccga gtgagagaca caaaaaattc caacacacta ttgcaatgaa
     6301 aataaatttc ctttattagc cagaagtcag atgctcaagg ggcttcatga tgtccccata
     6361 atttttggca gagggaaaaa gatctcagtg gtatttgtga gccagggcat tggccacacc
     6421 agccaccacc ttctgatagg cagcctgcac ctgaggagtg aattcacgac aggccttcga
     6481 atcaggcacc gggcttgcgg gtcatgcacc aggtgcgcgg tccttcgggc acctcgacgt
     6541 cggcggtgac ggtgaagccg agccgctcgt agaaggggag gttgcggggc gcggaggtct
     6601 ccaggaaggc gggcaccccg gcgcgctcgg ccgcctccac tccggggagc acgacggcgc
     6661 tgcccagacc cttgccctgg tggtcgggcg agacgccgac ggtggccagg aaccacgcgg
     6721 gctccttggg ccggtgcggc gccaggaggc cttccatctg ttgctgcgcg gccagccggg
     6781 aaccgctcaa ctcggccatg cgcgggccga tctcggcgaa caccgccccc gcttcgacgc
     6841 tctccggcgt ggtccagacc gccaccgcgg cgccgtcgtc cgcgacccac accttgccga
     6901 tgtcgagccc gacgcgcgtg aggaagagtt cttgcagctc ggtgacccgc tcgatgtggc
     6961 ggtccggatc gacggtgtgg cgcgtggcgg ggtagtcggc gaacgcggcg gcgagggtgc
     7021 gtacggccct ggggacgtcg tcgcgggtgg cgaggcgcac cgtgggcttg tactcggtca
     7081 tggtggcctc aggtgcagag gtttctacag ggaaagagag aaaggctaca gttagtttgc
     7141 tgtgatgatg ctaagtgccg aggtcagatg tctatggatt gtacacggaa tcccaacgca
     7201 ggaaccaaaa aacactcacc caaaaaaatg gcgctaaggc cgagagaaag gcgaagttcc
     7261 gtctacggct atataccttg aaacaggggc ggcagaagtg acgcaacgcg gttgcgcgga
     7321 cggcttcctg tgttttcatt ggtcgaagcg ttctgcgcgt gcgcagagat tccccaggga
     7381 aggcgggccc tcttccgaat gtccaatccg gacattcttc cggttggacc tccgcggtca
     7441 gcgctgtgcc cggcgagaga tcacgtgggg cgcggaggcg gtgctgctgg ggcacggccg
     7501 tccagcctcg gcggccatat ttttgagggg ctgttcatct cgttcacacg ctctgtccgc
     7561 catgtttgtg agtggaagcg ccattacccc ttcaagcgac tgaaggctgc agggcctctg
     7621 gtggcccgca tggggagacc agacccgcca ggcccgcctt tccgcactca gtccgggctt
     7681 actttatttt gtgagacagg gtctcgccta gag
//
