LOCUS       sequence-346555-        5655 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TEV (C-lobe) fused with FRB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5655)
  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 5655)
  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-TEV Component 2
FEATURES             Location/Qualifiers
     source          1..5655
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_recomb     167..994
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             167..291
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(329..934)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     enhancer        1062..1365
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1367..1569
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1520..1540
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1772..1794
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1856..1875
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     1917..1936
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1917..1935
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1936..1960
                     /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        1976..2085
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     CDS             2103..2132
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2163..2438
                     /codon_start=1
                     /product="T2098L mutant of FKBP-rapamycin binding domain of
                     human FRAP"
                     /label=FRB*
                     /note="binds synthetic ligands such as AP21967"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRIS"
     CDS             2823..2846
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(2936..2955)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3001..3056
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3055..3074)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     terminator      3144..3191
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     3204..3909
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           3292..3664
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(3321..3909)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     protein_bind    complement(3918..3951)
                     /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(4119..4706)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4198..4217)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4719..5510,5511..5579))
                     /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     5342..5361
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 atcaaatccc aagatgtgta taaacgcgcc ggtatgtaca ggaagaggtt tatactaaac
       61 tgttacattg caaacgtggt ttcgtgtgcc aagtgtgaaa accgatgttt aatcaaggct
      121 ctgacgcatt tctacaacca cgactccaag tgtgtgggtg aagtcatgca tcttttaatc
      181 aaatcccaag atgtgtataa accaccaaac tgccaaaaaa tgaaaactgt cgacaagctc
      241 tgtccgtttg ctggcaactg caagggtctc aatcctattt gtaattattg aataataaaa
      301 caattataaa tgtcaaattt gttttttatt aacgatacaa accaaacgca acaagaacat
      361 ttgtagtatt atctataatt gaaaacgcgt agttataatc gctgaggtaa tatttaaaat
      421 cattttcaaa tgattcacag ttaatttgcg acaatataat tttattttca cataaactag
      481 acgccttgtc gtcttcttct tcgtattcct tctctttttc atttttctct tcataaaaat
      541 taacatagtt attatcgtat ccatatatgt atctatcgta tagagtaaat tttttgttgt
      601 cataaatata tatgtctttt ttaatggggt gtatagtacc gctgcgcata gtttttctgt
      661 aatttacaac agtgctattt tctggtagtt cttcggagtg tgttgcttta attattaaat
      721 ttatataatc aatgaatttg ggatcgtcgg ttttgtacaa tatgttgccg gcatagtacg
      781 cagcttcttc tagttcaatt acaccatttt ttagcagcac cggattaaca taactttcca
      841 aaatgttgta cgaaccgtta aacaaaaaca gttcacctcc cttttctata ctattgtctg
      901 cgagcagttg tttgttgtta aaaataacag ccattgtaat gagacgcaca aactaatatc
      961 acaaactgga aatgtctatc aatatatagt tgctgatggc cggcctatta atagtaatca
     1021 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     1081 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     1141 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     1201 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtccgcc ccctattgac
     1261 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt acgggacttt
     1321 cctacttggc agtacatcta cgtattagtc atcgctatta ccatgctgat gcggttttgg
     1381 cagtacacca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     1441 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     1501 aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     1561 agcagacgtc gtttagtgaa ccgtcagatc actagatgct ttattgcggt agtttatcac
     1621 agttaaattg ctaacgccag tctcgaactt aacgtgcaga agttggtcgt gaggcactgg
     1681 gcaggtaagt atcgggccct ttgtgcgggg ggagcggctc ggggctgtcc gcggggggac
     1741 ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc
     1801 tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac
     1861 gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tggatcggac cgaaattaat
     1921 acgactcact ataggggaat tgtgagcgga taacaattcc ccggagttaa tccgggacct
     1981 ttaattcaac ccaacacaat atattatagt taaataagaa ttattatcaa atcatttgta
     2041 tattaattaa aatactatac tgtaaattac attttattta caatcaaagg agatatacca
     2101 tggagcagaa gctgatctcc gaggaggacc tgggatctgg atccgaattc atggcttcta
     2161 gaatcctctg gcatgagatg tggcatgaag gcctggaaga ggcatctcgt ttgtactttg
     2221 gggaaaggaa cgtgaaaggc atgtttgagg tgctggagcc cttgcatgct atgatggaac
     2281 ggggccccca gactctgaag gaaacatcct ttaatcaggc ctatggtcga gatttaatgg
     2341 aggcccaaga gtggtgcagg aagtacatga aatcagggaa tgtcaaggac ctcctccaag
     2401 cctgggacct ctattatcat gtgttccgac gaatctcagg tggatcaggt ggcgcggccg
     2461 cgggaatgtc tagcatggtg tcagacacta gttgcacatt cccttcatct gatggtatat
     2521 tctggaagca ttggattcaa accaaggatg ggcagtgtgg cagtccatta gtatcaacta
     2581 gagatgggtt cattgttggt atacactcag catcgaattt caccaacaca aacaattatt
     2641 tcacaagcgt gccgaaaaac ttcatggaat tgttgacaaa tcaggaggcg cagcagtggg
     2701 ttagtggttg gcgattaaac gctgactcag tattgtgggg gggccataaa gttttcatgg
     2761 tgaaacctga agaacctttt cagccagtta aggaagcgac tcaactcatg aattgactcg
     2821 agcaccacca tcaccatcac catcactaag tgattaacct caggtgcagg ctgcctatca
     2881 gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag atcgatcttt
     2941 ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct
     3001 aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg
     3061 gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt
     3121 tggcaacata tgcccatatg taactagcat aaccccttgg ggcctctaaa cgggtcttga
     3181 ggggtttttt gctgaaagca tgcggaggaa attctccttg aagtttccct ggtgttcaaa
     3241 gtaaaggagt ttgcaccaga cgcacctctg ttcactggtc cggcgtatta aaacacgata
     3301 cattgttatt agtacattta ttaagcgcta gattctgtgc gttgttgatt tacagacaat
     3361 tgttgtacgt attttaataa ttcattaaat ttataatctt tagggtggta tgttagagcg
     3421 aaaatcaaat gattttcagc gtctttatat ctgaatttaa atattaaatc ctcaatagat
     3481 ttgtaaaata ggtttcgatt agtttcaaac aagggttgtt tttccgaacc gatggctgga
     3541 ctatctaatg gattttcgct caacgccaca aaacttgcca aatcttgtag cagcaatcta
     3601 gctttgtcga tattcgtttg tgttttgttt tgtaataaag gttcgacgtc gttcaaaata
     3661 ttatgcgctt ttgtatttct ttcatcactg tcgttagtgt acaattgact cgacgtaaac
     3721 acgttaaata gagcttggac atatttaaca tcgggcgtgt tagctttatt aggccgatta
     3781 tcgtcgtcgt cccaaccctc gtcgttagaa gttgcttccg aagacgattt tgccatagcc
     3841 acacgacgcc tattaattgt gtcggctaac acgtccgcga tcaaatttgt agttgagctt
     3901 tttggaattg cgatcgcata acttcgtata gcatacatta tacgaagtta taagctcgga
     3961 acgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4021 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4081 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgccccctg
     4141 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4201 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4261 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4321 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4381 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4441 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4501 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4561 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4621 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4681 ttacgcgcag aaaaaaagga tctcaagaag atcctttgtt accaatgctt aatcagtgag
     4741 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4801 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4861 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4921 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4981 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5041 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5101 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5161 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5221 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5281 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     5341 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     5401 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     5461 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     5521 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     5581 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gtccgtgcgt
     5641 ttcctgcatc tttta
//
