LOCUS       sequence-243591-        8067 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SB-transposon plasmid for stable expression of unphosphorylatable 
            ""dead"" version of lipid raft-targeted Lyn-AktAR2-EV variant of 
            FRET-based AKT activity reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8067)
  TITLE     Eric Davis SB transposon plasmids
REFERENCE   2  (bases 1 to 8067)
  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"
COMMENT     Sequence Label: pSBbi-Pur-Lyn-AktAR2-EV-D
FEATURES             Location/Qualifiers
     source          1..8067
                     /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             2879..3613
                     /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(3667..3684)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3673..3897
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        4329..4433
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4434..4502,4503..5294)
                     /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(4652..4671)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5465..6053
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5954..5973
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6207..6224
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6598..6617
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    6616..6671
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     6717..6736
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(6836..7435)
                     /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(6919..6939)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     7416..7435
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        join(8067,1..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 tcgacggaac
     2461 ctctgcagga ggttccgcag ggggttccgc cggaggaagc gccggtggct ctgctggcgg
     2521 ctctgctggt gggtctgccg gcggatctgc gggagggagt gcgggtggaa gcgcaggagg
     2581 cagtgccggg ggatcagcgg ggggaagtgc cgggggtagt gctggtggct ctgcaggagg
     2641 ttccgcaggg ggttccgccg gaggaagcgc cggtggctct gctggcggct ctgctggtgg
     2701 gtctgccggc ggatctgcgg gagggagtgc gggtggaagc gcaggaggca gtgccggggg
     2761 atcagcggga ggaagtgccg ggggtagtgc tggtggcagc ggcgtcgacc ctcgtccgcg
     2821 ctcgtgcgtg tggccggacc ccaggccgga gtttggagga accggcggca gcgagctcat
     2881 gggcggcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     2941 gctgctgccc gacaaccact acctgagcta ccagtccaag ctgagcaaag accccaacga
     3001 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     3061 ggacgagctg tacaagggag gtaccggtgg atctatggtg agcaagggcg aggagctgtt
     3121 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     3181 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctgatctg
     3241 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcct
     3301 tcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     3361 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     3421 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     3481 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     3541 caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg
     3601 ccacaacatc gagtaaggcc tgtcaggcca agcttgggga tcaattctct agagctcgct
     3661 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     3721 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     3781 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     3841 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     3901 ctgaggcgga aagaaccagc tgggcctact agatagcgga ccccttaccg aaacatcgcc
     3961 gcattctgca gaggagtcga acttagtgta tgtaaacttc tgacccactg gaattgtgat
     4021 acagtgaatt ataagtgaaa taatctgtct gtaaacaatt gttggaaaaa tgacttgtgt
     4081 catgcacaaa gtagatgtcc taactgactt gccaaaacta ttgtttgtta acaagaaatt
     4141 tgtggagtag ttgaaaaacg agttttaatg actccaactt aagtgtatgt aaacttccga
     4201 cttcaactgt atagggatcc aattttaaca aaatattaac gcttacaatt tcctgatgcg
     4261 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atcaggtggc acttttcggg
     4321 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4381 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4441 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4501 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4561 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4621 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4681 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4741 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4801 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4861 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4921 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4981 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5041 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5101 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5161 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5221 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5281 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     5341 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5401 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5461 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5521 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5581 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5641 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5701 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5761 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5821 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5881 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5941 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6001 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6061 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6121 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     6181 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gatccctata
     6241 cagttgaagt cggaagttta catacacctt agccaaatac atttaaactc actttttcac
     6301 aattcctgac atttaatcct agtaaaaatt ccctgtctta ggtcagttag gatcaccact
     6361 ttattttaag aatgtgaaat atcagaataa tagtagagag aatgattcat ttcagctttt
     6421 atttctttca tcacattccc agtgggtcag aagtttacat acactcgact cctctgcaga
     6481 atgcggcgat gtttcggtaa ggggtcccga gtagctagtt catggcagcc agcatatggc
     6541 atatgttgcc aaactctaaa ccaaatactc attctgatgt tttaaatgat ttgccctccc
     6601 atatgtcctt ccgagtgaga gacacaaaaa attccaacac actattgcaa tgaaaataaa
     6661 tttcctttat tagccagaag tcagatgctc aaggggcttc atgatgtccc cataattttt
     6721 ggcagaggga aaaagatctc agtggtattt gtgagccagg gcattggcca caccagccac
     6781 caccttctga taggcagcct gcacctgagg agtgaattca cgacaggcct tcgaatcagg
     6841 caccgggctt gcgggtcatg caccaggtgc gcggtccttc gggcacctcg acgtcggcgg
     6901 tgacggtgaa gccgagccgc tcgtagaagg ggaggttgcg gggcgcggag gtctccagga
     6961 aggcgggcac cccggcgcgc tcggccgcct ccactccggg gagcacgacg gcgctgccca
     7021 gacccttgcc ctggtggtcg ggcgagacgc cgacggtggc caggaaccac gcgggctcct
     7081 tgggccggtg cggcgccagg aggccttcca tctgttgctg cgcggccagc cgggaaccgc
     7141 tcaactcggc catgcgcggg ccgatctcgg cgaacaccgc ccccgcttcg acgctctccg
     7201 gcgtggtcca gaccgccacc gcggcgccgt cgtccgcgac ccacaccttg ccgatgtcga
     7261 gcccgacgcg cgtgaggaag agttcttgca gctcggtgac ccgctcgatg tggcggtccg
     7321 gatcgacggt gtggcgcgtg gcggggtagt cggcgaacgc ggcggcgagg gtgcgtacgg
     7381 ccctggggac gtcgtcgcgg gtggcgaggc gcaccgtggg cttgtactcg gtcatggtgg
     7441 cctcaggtgc agaggtttct acagggaaag agagaaaggc tacagttagt ttgctgtgat
     7501 gatgctaagt gccgaggtca gatgtctatg gattgtacac ggaatcccaa cgcaggaacc
     7561 aaaaaacact cacccaaaaa aatggcgcta aggccgagag aaaggcgaag ttccgtctac
     7621 ggctatatac cttgaaacag gggcggcaga agtgacgcaa cgcggttgcg cggacggctt
     7681 cctgtgtttt cattggtcga agcgttctgc gcgtgcgcag agattcccca gggaaggcgg
     7741 gccctcttcc gaatgtccaa tccggacatt cttccggttg gacctccgcg gtcagcgctg
     7801 tgcccggcga gagatcacgt ggggcgcgga ggcggtgctg ctggggcacg gccgtccagc
     7861 ctcggcggcc atatttttga ggggctgttc atctcgttca cacgctctgt ccgccatgtt
     7921 tgtgagtgga agcgccatta ccccttcaag cgactgaagg ctgcagggcc tctggtggcc
     7981 cgcatgggga gaccagaccc gccaggcccg cctttccgca ctcagtccgg gcttacttta
     8041 ttttgtgaga cagggtctcg cctagag
//
