LOCUS       sequence-314517-        7776 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Blue-light dependent heterodimerization system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7776)
  AUTHORS   Benedetti L, Marvin JS, Falahati H, Guillen-Samander A, Looger LL, 
            De Camilli P
  TITLE     Optimized Vivid-derived Magnets photodimerizers for subcellular 
            optogenetics in mammalian cells.
  JOURNAL   Elife. 2020 Nov 11;9. pii: 63230. doi: 10.7554/eLife.63230.
  PUBMED    33174843
REFERENCE   2  (bases 1 to 7776)
  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: "Elife."; 
            date: "2020-11-11"; volume: "9. pii: 63230. doi"; pages: " 
            10.7554/eLife.63230"
COMMENT     Sequence Label: TagRFP-T-VAPB(1- 218)- eMagB
FEATURES             Location/Qualifiers
     source          1..7776
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      1772..1781
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1778..2509
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     intron          3700..3832
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3815..3834
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3841..4429
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3894..3914)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4312..4323)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4456..4473)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4462..4573
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4800..4819)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4865..4920
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4919..4938)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5279..5295)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5292..5314)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5303..5319
                     /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        complement(join(5327..5333,5334..5351,5352..5357))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5372..5393
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5452..5647
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5498..5633
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5653..5787
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5690..5709)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5744..5763
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5855..5872)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6026..6614)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6106..6125)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6785..7576,7577..7645))
                     /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     7408..7427
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7646..7750)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg ggcgagggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tgttgtgctg tctcatcatt ttggcaaaga attcggtccc gcgggcccgg
     1741 catccaccgg aagccacctg catgcggccg cgccaccatg gtgtctaagg gcgaagagct
     1801 gattaaggag aacatgcaca tgaagctgta catggagggc accgtgaaca accaccactt
     1861 caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca tgagaatcaa
     1921 ggtggtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacca gcttcatgta
     1981 cggcagcaga accttcatca accacaccca gggcatcccc gacttcttta agcagtcctt
     2041 ccctgagggc ttcacatggg agagagtcac cacatacgaa gacgggggcg tgctgaccgc
     2101 tacccaggac accagcctcc aggacggctg cctcatctac aacgtcaaga tcagaggggt
     2161 gaacttccca tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg aggccaacac
     2221 cgagatgctg taccccgctg acggcggcct ggaaggcaga accgacatgg ccctgaagct
     2281 cgtgggcggg ggccacctga tctgcaactt caagaccaca tacagatcca agaaacccgc
     2341 taagaacctc aagatgcccg gcgtctacta tgtggaccac agactggaaa gaatcaagga
     2401 ggccgacaaa gagacctacg tcgagcagca cgaggtggct gtggccagat actgcgacct
     2461 ccctagcaaa ctggggcaca aacttaatgg catggacgag ctgtacaagg gtacccgagc
     2521 tcaagcttcg aattcgatgg cgaaggtgga gcaggtcctg agtctcgagc cgcagcacga
     2581 gctcaaattc cgaggtccct tcaccgatgt tgtcaccacc aacctaaagc ttggcaaccc
     2641 gacagaccga aatgtgtgtt ttaaggtgaa gactacagca ccacgtaggt actgtgtgag
     2701 gcccaacagc ggaatcatcg atgcaggggc ctcaattaat gtatctgtga tgttacagcc
     2761 tttcgattat gatcccaatg agaaaagtaa acacaagttt atggttcagt ctatgtttgc
     2821 tccaactgac acttcagata tggaagcagt atggaaggag gcaaaaccgg aagaccttat
     2881 ggattcaaaa cttagatgtg tgtttgaatt gccagcagag aatgataaac cacatgatgt
     2941 agaaataaat aaaattatat ccacaactgc atcaaagaca gaaacaccaa tagtgtctaa
     3001 gtctctgagt tcttctttgg atgacaccga agttaagaag gttatggaag aatgtaagag
     3061 gctgcaaggt gaagttcaga ggctacggga ggagaacaag cagttcaagg aagaagatgg
     3121 actgcggatg aggaagacag tgcagagcaa cagccccatt tcagcattag ccccaactgg
     3181 gaaggaagag atatcgatgg gacataccct ctacgcgccg gggggttatg acatcatggg
     3241 ttacctcaga cagatcagaa accggccgaa cccacaagtg gagctgggac ccgtcgacct
     3301 ctcctgcgcc ctcgtgctgt gtgaccttaa gcagaaggac acccctgtgg tgtacgcctc
     3361 cgaagcattc ctggagatga ccgggtacaa cagacacgaa gtgctgggac ggaactgccg
     3421 cttcctgcaa tccccggatg gaatggtgaa gcctaagtca acccgcaaat acgtggactc
     3481 caacactatc ttcaccatga agaaggccat tgaccgcaat gcagaggtgc aagtggaagt
     3541 ggtgaacttc aagaagaacg gacagcgctt cgtcaacttc ctgactatga ttcccgtgcg
     3601 ggacgaaacc ggcgaatacc ggtacagcat cgggtttcag tgcgagactg agtaagttaa
     3661 cctcgaggtc ccgatgaata gctaaggtcg aggccgcagg taagtatcaa ggttacaaga
     3721 caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc
     3781 tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac aggtgtcgac
     3841 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     3901 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     3961 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     4021 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     4081 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     4141 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     4201 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
     4261 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     4321 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     4381 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaattcga
     4441 gctcggtacg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     4501 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     4561 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     4621 aggacagcaa gggggaggat tgggaagaca atagcaggcc agcttttgtt ccctttaggg
     4681 ttaattgcgc gcttggcgct aatcatggtc atagctgttt ccactcctca ggtgcaggct
     4741 gcctatcaga aggtggtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat
     4801 ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct
     4861 ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca
     4921 ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag
     4981 agtttggcaa catatgccat atgctggctg ccatgaacaa aggtggctat aaagaggtca
     5041 tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg
     5101 aggttagatt tttttttata ttttgtttgt gttatttttt tctttaacat ccctaaaatt
     5161 ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc
     5221 tgtccctctt ctcttatgaa gatccctcga cctgcagccc aagcttggcg taatcatggt
     5281 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5341 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5401 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa
     5461 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     5521 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     5581 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     5641 ttgcaaaaag ctaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5701 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5761 atcaatgtat cttatcatgt ctggatccgc tgcattaatg aatcggccaa cgcgcgggga
     5821 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5881 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5941 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6001 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6061 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6121 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6181 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
     6241 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6301 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6361 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6421 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     6481 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6541 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6601 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6661 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6721 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6781 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6841 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6901 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6961 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     7021 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     7081 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     7141 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7201 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7261 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7321 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7381 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7441 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7501 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7561 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7621 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7681 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7741 gggttccgcg cacatttccc cgaaaagtgc cacctg
//
