LOCUS       sequence-243607-        8031 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SB-transposon plasmid for stable expression of lipid raft-targeted 
            FRET-based kinase activity reporter with EV spacer, with AarI 
            cassette for cloning of a kinase-target peptide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8031)
  TITLE     Eric Davis SB transposon plasmids
REFERENCE   2  (bases 1 to 8031)
  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-LPAR
FEATURES             Location/Qualifiers
     source          1..8031
                     /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             2843..3577
                     /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(3631..3648)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3637..3861
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        4293..4397
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4398..4466,4467..5258)
                     /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(4616..4635)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5429..6017
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5918..5937
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6171..6188
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6562..6581
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    6580..6635
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     6681..6700
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(6800..7399)
                     /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(6883..6903)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     7380..7399
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        join(8031,1..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 ggcgtcgaca gtagcaggtg
     2821 tgaatcacct gctactgagc tcatgggcgg cgtgcagctc gccgaccact accagcagaa
     2881 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc
     2941 caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     3001 cgccgccggg atcactctcg gcatggacga gctgtacaag ggaggtaccg gtggatctat
     3061 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     3121 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     3181 caagctgacc ctgaagctga tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     3241 cgtgaccacc ctgggctacg gccttcagtg cttcgcccgc taccccgacc acatgaagca
     3301 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     3361 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     3421 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     3481 gctggagtac aactacaaca gccacaacgt ctatatcacc gccgacaagc agaagaacgg
     3541 catcaaggcc aacttcaaga tccgccacaa catcgagtaa ggcctgtcag gccaagcttg
     3601 gggatcaatt ctctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat
     3661 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     3721 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     3781 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     3841 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctgggcc tactagatag
     3901 cggacccctt accgaaacat cgccgcattc tgcagaggag tcgaacttag tgtatgtaaa
     3961 cttctgaccc actggaattg tgatacagtg aattataagt gaaataatct gtctgtaaac
     4021 aattgttgga aaaatgactt gtgtcatgca caaagtagat gtcctaactg acttgccaaa
     4081 actattgttt gttaacaaga aatttgtgga gtagttgaaa aacgagtttt aatgactcca
     4141 acttaagtgt atgtaaactt ccgacttcaa ctgtataggg atccaatttt aacaaaatat
     4201 taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4261 ccgcatcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4321 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4381 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4441 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4501 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4561 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     4621 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     4681 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4741 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4801 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4861 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     4921 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     4981 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5041 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5101 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5161 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5221 tcgctgaaat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5281 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5341 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5401 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5461 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5521 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5581 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5641 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5701 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5761 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5821 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5881 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5941 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6001 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6061 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6121 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     6181 tgagcgagga agcggatccc tatacagttg aagtcggaag tttacataca ccttagccaa
     6241 atacatttaa actcactttt tcacaattcc tgacatttaa tcctagtaaa aattccctgt
     6301 cttaggtcag ttaggatcac cactttattt taagaatgtg aaatatcaga ataatagtag
     6361 agagaatgat tcatttcagc ttttatttct ttcatcacat tcccagtggg tcagaagttt
     6421 acatacactc gactcctctg cagaatgcgg cgatgtttcg gtaaggggtc ccgagtagct
     6481 agttcatggc agccagcata tggcatatgt tgccaaactc taaaccaaat actcattctg
     6541 atgttttaaa tgatttgccc tcccatatgt ccttccgagt gagagacaca aaaaattcca
     6601 acacactatt gcaatgaaaa taaatttcct ttattagcca gaagtcagat gctcaagggg
     6661 cttcatgatg tccccataat ttttggcaga gggaaaaaga tctcagtggt atttgtgagc
     6721 cagggcattg gccacaccag ccaccacctt ctgataggca gcctgcacct gaggagtgaa
     6781 ttcacgacag gccttcgaat caggcaccgg gcttgcgggt catgcaccag gtgcgcggtc
     6841 cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag aaggggaggt
     6901 tgcggggcgc ggaggtctcc aggaaggcgg gcaccccggc gcgctcggcc gcctccactc
     6961 cggggagcac gacggcgctg cccagaccct tgccctggtg gtcgggcgag acgccgacgg
     7021 tggccaggaa ccacgcgggc tccttgggcc ggtgcggcgc caggaggcct tccatctgtt
     7081 gctgcgcggc cagccgggaa ccgctcaact cggccatgcg cgggccgatc tcggcgaaca
     7141 ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg ccgtcgtccg
     7201 cgacccacac cttgccgatg tcgagcccga cgcgcgtgag gaagagttct tgcagctcgg
     7261 tgacccgctc gatgtggcgg tccggatcga cggtgtggcg cgtggcgggg tagtcggcga
     7321 acgcggcggc gagggtgcgt acggccctgg ggacgtcgtc gcgggtggcg aggcgcaccg
     7381 tgggcttgta ctcggtcatg gtggcctcag gtgcagaggt ttctacaggg aaagagagaa
     7441 aggctacagt tagtttgctg tgatgatgct aagtgccgag gtcagatgtc tatggattgt
     7501 acacggaatc ccaacgcagg aaccaaaaaa cactcaccca aaaaaatggc gctaaggccg
     7561 agagaaaggc gaagttccgt ctacggctat ataccttgaa acaggggcgg cagaagtgac
     7621 gcaacgcggt tgcgcggacg gcttcctgtg ttttcattgg tcgaagcgtt ctgcgcgtgc
     7681 gcagagattc cccagggaag gcgggccctc ttccgaatgt ccaatccgga cattcttccg
     7741 gttggacctc cgcggtcagc gctgtgcccg gcgagagatc acgtggggcg cggaggcggt
     7801 gctgctgggg cacggccgtc cagcctcggc ggccatattt ttgaggggct gttcatctcg
     7861 ttcacacgct ctgtccgcca tgtttgtgag tggaagcgcc attacccctt caagcgactg
     7921 aaggctgcag ggcctctggt ggcccgcatg gggagaccag acccgccagg cccgcctttc
     7981 cgcactcagt ccgggcttac tttattttgt gagacagggt ctcgcctaga g
//
