LOCUS       sequence-243604-        8046 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SB-transposon plasmid for stable expression of lipid raft-targeted 
            FRET-based PKA activity reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8046)
  TITLE     Eric Davis SB transposon plasmids
REFERENCE   2  (bases 1 to 8046)
  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-Puro_LPAR_AKAR-WT
FEATURES             Location/Qualifiers
     source          1..8046
                     /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             2858..3592
                     /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(3646..3663)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3652..3876
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        4308..4412
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4413..4481,4482..5273)
                     /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(4631..4650)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5444..6032
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5933..5952
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6186..6203
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6577..6596
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    6595..6650
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     6696..6715
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(6815..7414)
                     /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(6898..6918)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     7395..7414
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        join(8046,1..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 caaagggcct
     2821 gcggagagct acactcgtgg atggcggaac agagctcatg ggcggcgtgc agctcgccga
     2881 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     2941 cctgagctac cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     3001 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggagg
     3061 taccggtgga tctatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     3121 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     3181 cgatgccacc tacggcaagc tgaccctgaa gctgatctgc accaccggca agctgcccgt
     3241 gccctggccc accctcgtga ccaccctggg ctacggcctt cagtgcttcg cccgctaccc
     3301 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     3361 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     3421 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     3481 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcaccgccga
     3541 caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg agtaaggcct
     3601 gtcaggccaa gcttggggat caattctcta gagctcgctg atcagcctcg actgtgcctt
     3661 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     3721 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     3781 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     3841 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     3901 gggcctacta gatagcggac cccttaccga aacatcgccg cattctgcag aggagtcgaa
     3961 cttagtgtat gtaaacttct gacccactgg aattgtgata cagtgaatta taagtgaaat
     4021 aatctgtctg taaacaattg ttggaaaaat gacttgtgtc atgcacaaag tagatgtcct
     4081 aactgacttg ccaaaactat tgtttgttaa caagaaattt gtggagtagt tgaaaaacga
     4141 gttttaatga ctccaactta agtgtatgta aacttccgac ttcaactgta tagggatcca
     4201 attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct
     4261 gtgcggtatt tcacaccgca tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4321 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4381 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4441 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4501 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     4561 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     4621 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     4681 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     4741 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4801 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     4861 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4921 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     4981 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5041 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5101 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5161 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5221 aacgaaatag acagatcgct gaaataggtg cctcactgat taagcattgg taactgtcag
     5281 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5341 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5401 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5461 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5521 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5581 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5641 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5701 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5761 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5821 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5881 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5941 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6001 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6061 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     6121 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     6181 agcgcagcga gtcagtgagc gaggaagcgg atccctatac agttgaagtc ggaagtttac
     6241 atacacctta gccaaataca tttaaactca ctttttcaca attcctgaca tttaatccta
     6301 gtaaaaattc cctgtcttag gtcagttagg atcaccactt tattttaaga atgtgaaata
     6361 tcagaataat agtagagaga atgattcatt tcagctttta tttctttcat cacattccca
     6421 gtgggtcaga agtttacata cactcgactc ctctgcagaa tgcggcgatg tttcggtaag
     6481 gggtcccgag tagctagttc atggcagcca gcatatggca tatgttgcca aactctaaac
     6541 caaatactca ttctgatgtt ttaaatgatt tgccctccca tatgtccttc cgagtgagag
     6601 acacaaaaaa ttccaacaca ctattgcaat gaaaataaat ttcctttatt agccagaagt
     6661 cagatgctca aggggcttca tgatgtcccc ataatttttg gcagagggaa aaagatctca
     6721 gtggtatttg tgagccaggg cattggccac accagccacc accttctgat aggcagcctg
     6781 cacctgagga gtgaattcac gacaggcctt cgaatcaggc accgggcttg cgggtcatgc
     6841 accaggtgcg cggtccttcg ggcacctcga cgtcggcggt gacggtgaag ccgagccgct
     6901 cgtagaaggg gaggttgcgg ggcgcggagg tctccaggaa ggcgggcacc ccggcgcgct
     6961 cggccgcctc cactccgggg agcacgacgg cgctgcccag acccttgccc tggtggtcgg
     7021 gcgagacgcc gacggtggcc aggaaccacg cgggctcctt gggccggtgc ggcgccagga
     7081 ggccttccat ctgttgctgc gcggccagcc gggaaccgct caactcggcc atgcgcgggc
     7141 cgatctcggc gaacaccgcc cccgcttcga cgctctccgg cgtggtccag accgccaccg
     7201 cggcgccgtc gtccgcgacc cacaccttgc cgatgtcgag cccgacgcgc gtgaggaaga
     7261 gttcttgcag ctcggtgacc cgctcgatgt ggcggtccgg atcgacggtg tggcgcgtgg
     7321 cggggtagtc ggcgaacgcg gcggcgaggg tgcgtacggc cctggggacg tcgtcgcggg
     7381 tggcgaggcg caccgtgggc ttgtactcgg tcatggtggc ctcaggtgca gaggtttcta
     7441 cagggaaaga gagaaaggct acagttagtt tgctgtgatg atgctaagtg ccgaggtcag
     7501 atgtctatgg attgtacacg gaatcccaac gcaggaacca aaaaacactc acccaaaaaa
     7561 atggcgctaa ggccgagaga aaggcgaagt tccgtctacg gctatatacc ttgaaacagg
     7621 ggcggcagaa gtgacgcaac gcggttgcgc ggacggcttc ctgtgttttc attggtcgaa
     7681 gcgttctgcg cgtgcgcaga gattccccag ggaaggcggg ccctcttccg aatgtccaat
     7741 ccggacattc ttccggttgg acctccgcgg tcagcgctgt gcccggcgag agatcacgtg
     7801 gggcgcggag gcggtgctgc tggggcacgg ccgtccagcc tcggcggcca tatttttgag
     7861 gggctgttca tctcgttcac acgctctgtc cgccatgttt gtgagtggaa gcgccattac
     7921 cccttcaagc gactgaaggc tgcagggcct ctggtggccc gcatggggag accagacccg
     7981 ccaggcccgc ctttccgcac tcagtccggg cttactttat tttgtgagac agggtctcgc
     8041 ctagag
//
