LOCUS       sequence-173557-        8540 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses chimeric fusion of PH domain of Btk and kinase domain of 
            Akt1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8540)
  AUTHORS   Ebner M, Lucic I, Leonard TA, Yudushkin I
  TITLE     PI(3,4,5)P3 Engagement Restricts Akt Activity to Cellular Membranes.
  JOURNAL   Mol Cell. 2017 Feb 2;65(3):416-431.e6. doi: 
            10.1016/j.molcel.2016.12.028.
  PUBMED    28157504
REFERENCE   2  (bases 1 to 8540)
  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: 
            "10.1016/j.molcel.2016.12"; journalName: "Mol Cell"; date: 
            "2017-02-2- 2"; volume: "65"; issue: "3"; pages: "416-431.e6"
COMMENT     Sequence Label: pFBD-Btk(PH)Akt1(KD)
FEATURES             Location/Qualifiers
     source          1..8540
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      40..628
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     529..548
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     782..799
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  933..1157
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1224..1757)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1946..1951,1952..1968,1969..1974))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    2482..2530
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2555..2574
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(4319..4428)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        4452..4543
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4468..4485
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4592..4621
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             4628..4651
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4661..4681
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQA"
     polyA_signal    6452..6586
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6489..6508)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6543..6562
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  6615..6780
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      6964..7418
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7051..7070)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7260..7281
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7444..7548
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7549..7617,7618..8409)
                     /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(7767..7786)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
       61 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
      121 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
      181 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
      241 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
      301 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
      361 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
      421 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
      481 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
      541 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
      601 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
      661 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
      721 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
      781 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
      841 tctgtgcggt atttcacacc gcatagacca gccgcgtaac ctggcaaaat cggttacggt
      901 tgagtaataa atggatgccc tgcgtaagcg ggtgtgggcg gacaataaag tcttaaactg
      961 aacaaaatag atctaaacta tgacaataaa gtcttaaact agacagaata gttgtaaact
     1021 gaaatcagtc cagttatgct gtgaaaaagc atactggact tttgttatgg ctaaagcaaa
     1081 ctcttcattt tctgaagtgc aaattgcccg tcgtattaaa gaggggcgtg gccaagggca
     1141 tggtaaagac tatattcgcg gcgttgtgac aatttaccga acaactccgc ggccgggaag
     1201 ccgatctcgg cttgaacgaa ttgttaggtg gcggtacttg ggtcgatatc aaagtgcatc
     1261 acttcttccc gtatgcccaa ctttgtatag agagccactg cgggatcgtc accgtaatct
     1321 gcttgcacgt agatcacata agcaccaagc gcgttggcct catgcttgag gagattgatg
     1381 agcgcggtgg caatgccctg cctccggtgc tcgccggaga ctgcgagatc atagatatag
     1441 atctcactac gcggctgctc aaacttgggc agaacgtaag ccgcgagagc gccaacaacc
     1501 gcttcttggt cgaaggcagc aagcgcgatg aatgtcttac tacggagcaa gttcccgagg
     1561 taatcggagt ccggctgatg ttgggagtag gtggctacgt ctccgaactc acgaccgaaa
     1621 agatcaagag cagcccgcat ggatttgact tggtcagggc cgagcctaca tgtgcgaatg
     1681 atgcccatac ttgagccacc taactttgtt ttagggcgac tgccctgctg cgtaacatcg
     1741 ttgctgctgc gtaacatcgt tgctgctcca taacatcaaa catcgaccca cggcgtaacg
     1801 cgcttgctgc ttggatgccc gaggcataga ctgtacaaaa aaacagtcat aacaagccat
     1861 gaaaaccgcc actgcgccgt taccaccgct gcgttcggtc aaggttctgg accagttgcg
     1921 tgagcgcata cgctacttgc attacagttt acgaaccgaa caggcttatg tcaactgggt
     1981 tcgtgccttc atccgtttcc acggtgtgcg tcacccggca accttgggca gcagcgaagt
     2041 cgaggcattt ctgtcctggc tggcgaacga gcgcaaggtt tcggtctcca cgcatcgtca
     2101 ggcattggcg gccttgctgt tcttctacgg caaggtgctg tgcacggatc tgcccttgct
     2161 tcaggagatc ggtagacctc ggccgtcgcg gcgcttgccg gtggtgctga ccccggatga
     2221 agtggttcgc atcctcggtt ttctggaagg cgagcatcgt ttgttcgccc aggactctag
     2281 ctatagttct agtggttggc tacgtacccg tagtggctat ggcagggctt gccgccccga
     2341 cgttggctgc gagccctggg ccttcacccg aacttggggg ttggggtggg gaaaaggaag
     2401 aaacgcgggc gtattggtcc caatggggtc tcggtggggt atcgacagag tgccagccct
     2461 gggaccgaac cccgcgttta tgaacaaacg acccaacacc cgtgcgtttt attctgtctt
     2521 tttattgccg tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt ttcagttagc
     2581 ctcccccatc tcccggtacc tcactgcaca gcggcgtccg ggtggctctg gtatcgctgc
     2641 ctccaaacct cctggatctt cctgcaccac ttgtgtgcgt tcccgctggg gtccatcaga
     2701 taatacgtcc tgttaggcgt gtggacaaag aaagttttaa aattcttggc ctctggtcga
     2761 agttcttgtg accaaggaat ttcacctttc agaactttgt tgacaggatc cacataatat
     2821 aaatgtggtc cttctgtgag caacagctgt cgtcgtcttg caaataaacc cttccgctta
     2881 tccactgggc ccatctttag tattaaatta ttttctacaa actggtgcca agggtttccg
     2941 ccagcctgct tctccaacaa caacctcttc tcatcttcgg aaaactgtaa gtccagttca
     3001 aaggagttcg agtccagatc gtgaatgtac tgctctatgt tgctgcctga cctctggggc
     3061 aggcccgtgt cggaggctga cagggagtgt gaggaggagg acgaagacac ctgcatgcag
     3121 ccaaactggc tcaggagatt gtcataattg ccatagcagt cctcgtcgtc ttccgacata
     3181 gccggcaggt aagcggtgag cttcggaggc gtctgctggt gcaggttctc ccacgtgacg
     3241 gactcgaaga acgggtgtgc tttaagaggt ccgtatcctt ccatttcctc acagcctaac
     3301 cgctttgtgg catctaaaac caaaagtttc tccacgaggt ctcttgcctt agggaagaat
     3361 ttttctggaa agtcatattc caacttaatg atcttctgaa atataagata ctcgtttcca
     3421 gctcggaatg gtgggagtcc tgccacaagc tggtatatta tgcatccaag agcccaaagg
     3481 tctgaactct tacaggcgga cttctccgtg agcagctctg gagaaacgta ctgcgctgtt
     3541 cccacgaatg agttggccct ggcttgtttg ctctctgggg ataagacttt tgctgttcca
     3601 aaatctgtga tctggatgtg catatcttca tttaacaaaa tgttttccgg tttaaggtcc
     3661 ctgtgaatga tgcccttgcc gtgcaagtac tctaaagcag acacaatctc agccgtgtaa
     3721 aatcgggtac aggtctcatc gaatgaaccg attttgcgaa tatatttaag tagttctcca
     3781 tttttggcat aactaaggcc gaaatacagc ttctcgtcgt cctgaaatgt gaagtaaagc
     3841 ttaacaaaga aggggtgatc caggcgcgac atgacatccc gctctctggt tacatagggg
     3901 accttgttct cttttatgat atgtcgcttc tccagaattt taatcgcata ttctctggag
     3961 gttgccagtt ctcgagccag gacaaccgtg gaaaaagagc cttccccaag gattttccca
     4021 aacttgaagt cctcaggccg cttcttccga ggctgcggcg gaggctgggc atgctgcagg
     4081 gagccggcgc cgggccgagg ctcggctgca gtgccgtcca tggcggggcc ctgcctgctg
     4141 ccaccaggaa tgccaggggg cgtgctggac tcagtctggg tcctcaccat tgatggggat
     4201 gggcaggaac ataacaccac gctggactgg atgggcacgg cgtcatacag ctggctggtg
     4261 gtcctggcca tgctagcacc atggctcgag atcccgggtg atcaagtctt cgtcgagtga
     4321 ttgtaaataa aatgtaattt acagtatagt attttaatta atatacaaat gatttgataa
     4381 taattcttat ttaactataa tatattgtgt tgggttgaat taaaggtccg tatactccgg
     4441 aatattaata gatcatggag ataattaaaa tgataaccat ctcgcaaata aataagtatt
     4501 ttactgtttt cgtaacagtt ttgtaataaa aaaacctata aatattccgg attattcata
     4561 ccgtcccacc atcgggcgcg gatccatgtc gcaccatcac catcaccatc accatcacca
     4621 tgcaagctgg agccacccgc agttcgaaaa gggctccggt gaaaacctgt atttccaggc
     4681 gcgcatggcc gcagtgattc tggagagcat ctttctgaag cgatcccaac agaaaaagaa
     4741 aacatcacct ctaaacttca agaagcgcct gtttctcttg accgtgcaca aactctccta
     4801 ctatgagtat gactttgaac gtgggagaag aggcagtaag aagggttcaa tagatgttga
     4861 gaagatcact tgtgttgaaa cagtggttcc tgaaaaaaat cctcctccag aaagacagat
     4921 tccgagaaga ggtgaagagt ccagtgaaat ggagcaaatt tcaatcattg aaaggttccc
     4981 ttatcccttc caggttgtat atgatgaagg gcctctctac gtcttctccc caactgaaga
     5041 actaaggaag cggtggattc accagctcaa aaacgtaatc cggtacaaca gtgatctggt
     5101 tcagaaatat cacccttgct tctggatcga tgggcagtat ctctgctgct ctcagacagc
     5161 caaaaatgct atgggctgcc aaattttgga gaacatggac ttccggtcgg gctcacccag
     5221 tgacaactca ggggctgaag agatggaggt gtccctggcc aagcccaagc accgcgtgac
     5281 catgaacgag tttgagtacc tgaagctgct gggcaagggc actttcggca aggtgatcct
     5341 ggtgaaggag aaggccacag gccgctacta cgccatgaag atcctcaaga aggaagtcat
     5401 cgtggccaag gacgaggtgg cccacacact caccgagaac cgcgtcctgc agaactccag
     5461 gcaccccttc ctcacagccc tgaagtactc tttccagacc cacgaccgcc tctgctttgt
     5521 catggagtac gccaacgggg gcgagctgtt cttccacctg tcccgggagc gtgtgttctc
     5581 cgaggaccgg gcccgcttct atggcgctga gattgtgtca gccctggact acctgcactc
     5641 ggagaagaac gtggtgtacc gggacctcaa gctggagaac ctcatgctgg acaaggacgg
     5701 gcacattaag atcacagact tcgggctgtg caaggagggg atcaaggacg gtgccaccat
     5761 gaagaccttt tgcggcacac ctgagtacct ggcccccgag gtgctggagg acaatgacta
     5821 cggccgtgca gtggactggt gggggctggg cgtggtcatg tacgagatga tgtgcggtcg
     5881 cctgcccttc tacaaccagg accatgagaa gctttttgag ctcatcctca tggaggagat
     5941 ccgcttcccg cgcacgcttg gtcccgaggc caagtccttg ctttcagggc tgctcaagaa
     6001 ggaccccaag cagaggcttg gcgggggctc cgaggacgcc aaggagatca tgcagcatcg
     6061 cttctttgcc ggtatcgtgt ggcagcacgt gtacgagaag aagctcagcc cacccttcaa
     6121 gccccaggtc acgtcggaga ctgacaccag gtattttgat gaggagttca cggcccagat
     6181 gatcaccatc acaccacctg accaagatga cagcatggag tgtgtggaca gcgagcgcag
     6241 gccccacttc ccccagttct cctactcggc cagcagcacg gcctgagcgg ccgctttcga
     6301 atctagagcc tgcagtctcg acaagcttgt cgagaagtac tagaggatca taatcagcca
     6361 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     6421 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     6481 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6541 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatctgat cactgcttga
     6601 gcctaggaga tccgaaccag ataagtgaaa tctagttcca aactattttg tcatttttaa
     6661 ttttcgtatt agcttacgac gctacaccca gttcccatct attttgtcac tcttccctaa
     6721 ataatcctta aaaactccat ttccacccct cccagttccc aactattttg tccgcccaca
     6781 gcggggcatt tttcttcctg ttatgttttt aatcaaacat cctgccaact ccatgtgaca
     6841 aaccgtcatc ttcggctact ttttctctgt cacagaatga aaatttttct gtcatctctt
     6901 cgttattaat gtttgtaatt gactgaatat caacgcttat ttgcagcctg aatggcgaat
     6961 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     7021 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     7081 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     7141 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     7201 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttcttaatag
     7261 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     7321 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     7381 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     7441 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     7501 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     7561 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     7621 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     7681 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     7741 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     7801 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     7861 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     7921 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     7981 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     8041 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     8101 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     8161 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     8221 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     8281 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     8341 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     8401 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     8461 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     8521 taacgtgagt tttcgttcca
//
