LOCUS       sequence-347221-        7134 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  light-controlled analog of the alpha-tubulin acetylase alpha-TAT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7134)
  AUTHORS   Stone OJ, Pankow N, Liu B, Sharma VP, Eddy RJ, Wang H, Putz AT, 
            Teets FD, Kuhlman B, Condeelis JS, Hahn KM
  TITLE     Optogenetic control of cofilin and alphaTAT in living cells using 
            Z-lock.
  JOURNAL   Nat Chem Biol. 2019 Dec;15(12):1183-1190. doi: 
            10.1038/s41589-019-0405-4. Epub 2019 Nov 18.
  PUBMED    31740825
REFERENCE   2  (bases 1 to 7134)
  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: 
            "10.1038/s41589-019-0405-4"; journalName: "Nat Chem Biol"; date: 
            "2019-12"; volume: "15"; issue: "12"; pages: "1183-1190"
COMMENT     Sequence Label: Z-lock alpha-TAT
FEATURES             Location/Qualifiers
     source          1..7134
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     238..260
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     322..341
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     383..402
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        383..401
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    402..426
                     /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        442..551
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     CDS             569..592
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             605..1309
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(747..765)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1015..1034
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1322..1495
                     /codon_start=1
                     /product="""Zdark"" protein, derived from the Z subunit of 
                     protein A, that binds to the dark state of LOV2 and is 
                     released upon irradiation (Wang et al., 2016)"
                     /label=Zdk1
                     /translation="VDNKFNKEKTRAGAEIHSLPNLNVEQKFAFIVSLFDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             2261..2689
                     /codon_start=1
                     /gene="NPH1 from Avena sativa"
                     /product="light, oxygen, or voltage domain from phototropin
                     1 of Avena sativa (Herrou and Crosson, 2011)"
                     /label=LOV2
                     /note="photoswitch that undergoes a light-inducible 
                     conformational change"
                     /translation="LATTLERIEKNFVITDPRLPDNPIIFASDSFLQLTEYSREEILGR
                     NCRFLQGPETDRATVRKIRDAIDNQTEVTVQLINYTKSGKKFWNLFHLQPMRDQKGDVQ
                     YFIGVQLDGTEHVRDAAEREGVMLIKKTAENIDEAAKEL"
     CDS             2729..2761
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /label=HSV tag
                     /translation="QPELAPEDPED"
     CDS             2768..2791
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(2881..2900)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2946..3001
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3000..3019)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     terminator      3089..3136
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     3149..3854
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           3237..3609
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(3266..3854)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     protein_bind    complement(3863..3896)
                     /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(4064..4651)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4143..4162)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4664..5455,5456..5524))
                     /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     5287..5306
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     misc_recomb     5767..6594
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             5767..5891
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5929..6534)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     enhancer        6662..6965
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(6967..7134,1..35)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     join(7120..7134,1..6)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 ggcgtgtacg gtgggaggtc tatataagca gacgtcgttt agtgaaccgt cagatcacta
       61 gatgctttat tgcggtagtt tatcacagtt aaattgctaa cgccagtctc gaacttaacg
      121 tgcagaagtt ggtcgtgagg cactgggcag gtaagtatcg ggccctttgt gcggggggag
      181 cggctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt
      241 tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt catgccttct
      301 tctttttcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat cattttggca
      361 aagaattgga tcggaccgaa attaatacga ctcactatag gggaattgtg agcggataac
      421 aattccccgg agttaatccg ggacctttaa ttcaacccaa cacaatatat tatagttaaa
      481 taagaattat tatcaaatca tttgtatatt aattaaaata ctatactgta aattacattt
      541 tatttacaat caaaggagat ataccatgga ctataaggac gatgatgaca aaggatctgg
      601 atctgtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
      661 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
      721 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
      781 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
      841 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
      901 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
      961 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     1021 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     1081 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     1141 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     1201 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     1261 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagg gttctggatc
     1321 cgtggataac aaattcaata aagaaaagac gcgtgccggt gcggaaatcc attctctgcc
     1381 aaacctgaat gttgagcaga agtttgcctt catcgtgagc ctgtttgatg atccatctca
     1441 gagcgccaat ctgctggccg aagccaaaaa actgaacgat gcccaggccc caaaaggttc
     1501 tggaagtggt tcgggctcag gaagtgaatt cgagttcccg ttcgatgtgg atgcgctgtt
     1561 cccggagcgg atcaccgtgc tggaccagca cctgcggcct ccggcccgcc gacccggaac
     1621 cacaacgccg gcccgtgtgg atctgcagca gcaaatcatg actattgtag atgagctggg
     1681 caaggcttct gccaaggcgc agcacctccc tgcacccatc accagcgctt tgaggatgca
     1741 aagcaaccgg cacgttattt acatactgaa ggacacctca gcccgaccgg cagggaaagg
     1801 agccattatt ggtttcctca aagttggata caagaagctc tttgtactgg atgaccggga
     1861 ggctcacaat gaggtagaac ccctttgcat tctggacttt tacatccacg agtcggtgca
     1921 acggcatggc cacgggcgag aactttttca gcatatgtta cagaaagagc gagtggagcc
     1981 ccaccaactg gccattgacc gaccatcgcc gaagctgctc aagttcctga ataagcacta
     2041 caacctggag accacagtcc cacaggtgaa caactttgtc atctttgaag gcttctttgc
     2101 ccatcagcac cggcccccca cttcctctct gagagcaact cgacactctc gtgctgctgt
     2161 ggccgatccc atacctgctg ctccagcacg gaagctgcca ccaaaaagag cagagggaga
     2221 cattaagcca tactctggta ccggctcagg aagtggtagc ttggctacta cacttgaacg
     2281 tattgagaag aactttgtca ttactgaccc aagattgcca gataatccca ttatattcgc
     2341 gtccgatagt ttcttgcagt tgacagaata tagccgtgaa gaaattttgg gaagaaactg
     2401 caggtttcta caaggtcctg aaactgatcg cgcgacagtg agaaaaatta gagatgccat
     2461 agataaccaa acagaggtca ctgttcagct gattaattat acaaagagtg gtaaaaagtt
     2521 ctggaacctc tttcacttgc agcctatgcg agatcagaag ggagatgtcc agtactttat
     2581 tggggttcag ttggatggaa ctgagcatgt ccgagatgct gccgagagag agggagtcat
     2641 gctgattaag aaaactgcag aaaatattga tgaggcggca aaagaacttt aaaagcttgc
     2701 ggccgcacag ctgtatacac gtgcaagcca gccagaactc gccccggaag accccgagga
     2761 tctcgagcac caccatcacc atcaccatca ctaagtgatt aacctcaggt gcaggctgcc
     2821 tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatcga
     2881 tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc
     2941 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
     3001 actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta
     3061 gagtttggca acatatgccc atatgtaact agcataaccc cttggggcct ctaaacgggt
     3121 cttgaggggt tttttgctga aagcatgcgg aggaaattct ccttgaagtt tccctggtgt
     3181 tcaaagtaaa ggagtttgca ccagacgcac ctctgttcac tggtccggcg tattaaaaca
     3241 cgatacattg ttattagtac atttattaag cgctagattc tgtgcgttgt tgatttacag
     3301 acaattgttg tacgtatttt aataattcat taaatttata atctttaggg tggtatgtta
     3361 gagcgaaaat caaatgattt tcagcgtctt tatatctgaa tttaaatatt aaatcctcaa
     3421 tagatttgta aaataggttt cgattagttt caaacaaggg ttgtttttcc gaaccgatgg
     3481 ctggactatc taatggattt tcgctcaacg ccacaaaact tgccaaatct tgtagcagca
     3541 atctagcttt gtcgatattc gtttgtgttt tgttttgtaa taaaggttcg acgtcgttca
     3601 aaatattatg cgcttttgta tttctttcat cactgtcgtt agtgtacaat tgactcgacg
     3661 taaacacgtt aaatagagct tggacatatt taacatcggg cgtgttagct ttattaggcc
     3721 gattatcgtc gtcgtcccaa ccctcgtcgt tagaagttgc ttccgaagac gattttgcca
     3781 tagccacacg acgcctatta attgtgtcgg ctaacacgtc cgcgatcaaa tttgtagttg
     3841 agctttttgg aattgcgatc gcataacttc gtatagcata cattatacga agttataagc
     3901 tcggaacgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3961 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4021 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4081 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4141 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4201 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4261 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4321 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4381 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4441 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4501 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4561 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4621 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgttaccaa tgcttaatca
     4681 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4741 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4801 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4861 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4921 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4981 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5041 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5101 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5161 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5221 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5281 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5341 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5401 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5461 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5521 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgtccg
     5581 cgcgtttcct gcatctttta atcaaatccc aagatgtgta taaacgcgcc ggtatgtaca
     5641 ggaagaggtt tatactaaac tgttacattg caaacgtggt ttcgtgtgcc aagtgtgaaa
     5701 accgatgttt aatcaaggct ctgacgcatt tctacaacca cgactccaag tgtgtgggtg
     5761 aagtcatgca tcttttaatc aaatcccaag atgtgtataa accaccaaac tgccaaaaaa
     5821 tgaaaactgt cgacaagctc tgtccgtttg ctggcaactg caagggtctc aatcctattt
     5881 gtaattattg aataataaaa caattataaa tgtcaaattt gttttttatt aacgatacaa
     5941 accaaacgca acaagaacat ttgtagtatt atctataatt gaaaacgcgt agttataatc
     6001 gctgaggtaa tatttaaaat cattttcaaa tgattcacag ttaatttgcg acaatataat
     6061 tttattttca cataaactag acgccttgtc gtcttcttct tcgtattcct tctctttttc
     6121 atttttctct tcataaaaat taacatagtt attatcgtat ccatatatgt atctatcgta
     6181 tagagtaaat tttttgttgt cataaatata tatgtctttt ttaatggggt gtatagtacc
     6241 gctgcgcata gtttttctgt aatttacaac agtgctattt tctggtagtt cttcggagtg
     6301 tgttgcttta attattaaat ttatataatc aatgaatttg ggatcgtcgg ttttgtacaa
     6361 tatgttgccg gcatagtacg cagcttcttc tagttcaatt acaccatttt ttagcagcac
     6421 cggattaaca taactttcca aaatgttgta cgaaccgtta aacaaaaaca gttcacctcc
     6481 cttttctata ctattgtctg cgagcagttg tttgttgtta aaaataacag ccattgtaat
     6541 gagacgcaca aactaatatc acaaactgga aatgtctatc aatatatagt tgctgatggc
     6601 cggcctatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     6661 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     6721 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     6781 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     6841 caagtccgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     6901 acatgacctt acgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     6961 ccatgctgat gcggttttgg cagtacacca atgggcgtgg atagcggttt gactcacggg
     7021 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     7081 gggactttcc aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc ggta
//
