LOCUS       sequence-353998-        6117 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SPELL Lyn kinase (Constitutively active) N-lobe fused to iFKBP. 
            Split site is 279..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6117)
  AUTHORS   Dagliyan O, Krokhotin A, Ozkan-Dagliyan I, Deiters A, Der CJ, Hahn 
            KM, Dokholyan NV
  TITLE     Computational design of chemogenetic and optogenetic split proteins.
  JOURNAL   Nat Commun. 2018 Oct 2;9(1):4042. doi: 10.1038/s41467-018-06531-4.
  PUBMED    30279442
REFERENCE   2  (bases 1 to 6117)
  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"; journalName: "Nat 
            Commun."; date: "2018-10-2"; volume: "9(1):4042. doi"; pages: " 
            10.1038/s41467-018-06531-4"
COMMENT     Sequence Label: SPELL-Lyn Component 1
FEATURES             Location/Qualifiers
     source          1..6117
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_recomb     146..973
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             146..270
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(308..913)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     enhancer        1041..1344
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1346..1548
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1499..1519
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1751..1773
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1835..1854
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     1896..1915
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1896..1914
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1915..1939
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        1955..2064
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     CDS             2082..2105
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      2127..2136
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3264..3287
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(3377..3396)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3442..3497
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3496..3515)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     terminator      3585..3632
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     3645..4350
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           3733..4105
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(3762..4350)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     protein_bind    complement(4359..4392)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(4560..5147)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4639..4658)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5160..5951,5952..6020))
                     /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     5783..5802
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaacgcgccg gtatgtacag gaagaggttt atactaaact gttacattgc aaacgtggtt
       61 tcgtgtgcca agtgtgaaaa ccgatgttta atcaaggctc tgacgcattt ctacaaccac
      121 gactccaagt gtgtgggtga agtcatgcat cttttaatca aatcccaaga tgtgtataaa
      181 ccaccaaact gccaaaaaat gaaaactgtc gacaagctct gtccgtttgc tggcaactgc
      241 aagggtctca atcctatttg taattattga ataataaaac aattataaat gtcaaatttg
      301 ttttttatta acgatacaaa ccaaacgcaa caagaacatt tgtagtatta tctataattg
      361 aaaacgcgta gttataatcg ctgaggtaat atttaaaatc attttcaaat gattcacagt
      421 taatttgcga caatataatt ttattttcac ataaactaga cgccttgtcg tcttcttctt
      481 cgtattcctt ctctttttca tttttctctt cataaaaatt aacatagtta ttatcgtatc
      541 catatatgta tctatcgtat agagtaaatt ttttgttgtc ataaatatat atgtcttttt
      601 taatggggtg tatagtaccg ctgcgcatag tttttctgta atttacaaca gtgctatttt
      661 ctggtagttc ttcggagtgt gttgctttaa ttattaaatt tatataatca atgaatttgg
      721 gatcgtcggt tttgtacaat atgttgccgg catagtacgc agcttcttct agttcaatta
      781 caccattttt tagcagcacc ggattaacat aactttccaa aatgttgtac gaaccgttaa
      841 acaaaaacag ttcacctccc ttttctatac tattgtctgc gagcagttgt ttgttgttaa
      901 aaataacagc cattgtaatg agacgcacaa actaatatca caaactggaa atgtctatca
      961 atatatagtt gctgatggcc ggcctattaa tagtaatcaa ttacggggtc attagttcat
     1021 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     1081 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     1141 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     1201 catcaagtgt atcatatgcc aagtccgccc cctattgacg tcaatgacgg taaatggccc
     1261 gcctggcatt atgcccagta catgacctta cgggactttc ctacttggca gtacatctac
     1321 gtattagtca tcgctattac catgctgatg cggttttggc agtacaccaa tgggcgtgga
     1381 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     1441 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta ataaccccgc cccgttgacg
     1501 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagacgtcg tttagtgaac
     1561 cgtcagatca ctagatgctt tattgcggta gtttatcaca gttaaattgc taacgccagt
     1621 ctcgaactta acgtgcagaa gttggtcgtg aggcactggg caggtaagta tcgggccctt
     1681 tgtgcggggg gagcggctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg
     1741 ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat
     1801 gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct
     1861 catcattttg gcaaagaatt ggatcggacc gaaattaata cgactcacta taggggaatt
     1921 gtgagcggat aacaattccc cggagttaat ccgggacctt taattcaacc caacacaata
     1981 tattatagtt aaataagaat tattatcaaa tcatttgtat attaattaaa atactatact
     2041 gtaaattaca ttttatttac aatcaaagga gatataccat ggactataag gacgatgatg
     2101 acaaaggatc tggatccgaa ttcgccgcca ccatgggatg tataaaatca aaagggaaag
     2161 acagcttgag tgacgatgga gtagatttga agactcaacc agtacgtaat actgaaagaa
     2221 ctatttatgt gagagatcca acgtccaata aacagcaaag gccagttcca gaatctcagc
     2281 ttttacctgg acagaggttt caaactaaag atccagagga acaaggagac attgtggtag
     2341 ccttgtaccc ctatgatggc atccacccgg acgacttgtc tttcaagaaa ggagagaaga
     2401 tgaaagtcct ggaggagcat ggagaatggt ggaaagcaaa gtccctttta acaaaaaaag
     2461 aaggcttcat ccccagcaac tatgtggcca aactcaacac cttagaaaca gaagagtggt
     2521 ttttcaagga tataaccagg aaggacgcag aaaggcagct tttggcacca ggaaatagcg
     2581 ctggagcttt ccttattaga gaaagtgaaa cattaaaagg aagcttctct ctgtctgtca
     2641 gagactttga ccctgtgcat ggtgatgtta ttaagcacta caaaattaga agtctggata
     2701 atgggggcta ttacatctct ccacgaatca cttttccctg tatcagcgac atgattaaac
     2761 attaccaaaa gcaggcagat ggcttgtgca gaagattgga gaaggcttgt attagtccca
     2821 agccacagaa gccatgggat aaagatgcct gggagatccc ccgggagtcc atcaagttgg
     2881 tgaaaaggct tggcgctggg cagtttgggg aagtctggat gggttactat aacggaagta
     2941 ccaaggtggc tgtgaaaacc ctgaagccag gtggatcagg tggcgcggcc gcgacctgcg
     3001 tggtgcacta caccgggatg cttgaagatg gaaagaaatt tgattcctcc cgggacagaa
     3061 acaagccctt taagtttatg ctaggcaagc aggaggtgat ccgaggctgg gaagaagggg
     3121 ttgcccagat gagtgtgggt cagagagcca aactgactat atctccagat tatgcctatg
     3181 gtgccactgg gcacccaggc atcatcccac cacatgccac tctcgtcttc gatgtggagc
     3241 ttctaaaact ggaatgactc gagcaccacc atcaccatca ccatcactaa gtgattaacc
     3301 tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa
     3361 ataccactga gatcgatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
     3421 ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     3481 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga
     3541 atgagtattt ggtttagagt ttggcaacat atgcccatat gtaactagca taaccccttg
     3601 gggcctctaa acgggtcttg aggggttttt tgctgaaagc atgcggagga aattctcctt
     3661 gaagtttccc tggtgttcaa agtaaaggag tttgcaccag acgcacctct gttcactggt
     3721 ccggcgtatt aaaacacgat acattgttat tagtacattt attaagcgct agattctgtg
     3781 cgttgttgat ttacagacaa ttgttgtacg tattttaata attcattaaa tttataatct
     3841 ttagggtggt atgttagagc gaaaatcaaa tgattttcag cgtctttata tctgaattta
     3901 aatattaaat cctcaataga tttgtaaaat aggtttcgat tagtttcaaa caagggttgt
     3961 ttttccgaac cgatggctgg actatctaat ggattttcgc tcaacgccac aaaacttgcc
     4021 aaatcttgta gcagcaatct agctttgtcg atattcgttt gtgttttgtt ttgtaataaa
     4081 ggttcgacgt cgttcaaaat attatgcgct tttgtatttc tttcatcact gtcgttagtg
     4141 tacaattgac tcgacgtaaa cacgttaaat agagcttgga catatttaac atcgggcgtg
     4201 ttagctttat taggccgatt atcgtcgtcg tcccaaccct cgtcgttaga agttgcttcc
     4261 gaagacgatt ttgccatagc cacacgacgc ctattaattg tgtcggctaa cacgtccgcg
     4321 atcaaatttg tagttgagct ttttggaatt gcgatcgcat aacttcgtat agcatacatt
     4381 atacgaagtt ataagctcgg aacgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4441 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4501 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4561 ttccataggc tccgccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4621 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4681 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4741 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4801 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4861 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4921 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4981 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5041 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5101 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttgt
     5161 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5221 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5281 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5341 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5401 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5461 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5521 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5581 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5641 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5701 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5761 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5821 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5881 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccaac
     5941 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6001 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6061 tattgtctca tgtccgcgcg tttcctgcat cttttaatca aatcccaaga tgtgtat
//
