LOCUS       sequence-483128-        6927 bp    DNA     circular SYN 13-AUG-2025
DEFINITION  Blue light-induced reconstitution of mCherry and its relocalization 
            to the nucleus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6927)
  TITLE     Photoswitchable intein for light control of covalent protein binding
            and cleavage
REFERENCE   2  (bases 1 to 6927)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 13, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: PS Intein NLS-NES
FEATURES             Location/Qualifiers
     source          1..6927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(733..751)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1001..1020
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             join(1419..1421,1422..1424,1425..2135)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1464..1485)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     2072..2093
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2754..3203
                     /codon_start=1
                     /product="""positive Magnet"" variant of an N-terminally 
                     truncated Vivid (VVD) blue light photoreceptor from 
                     Neurospora crassa (Kawano et al., 2015)"
                     /label=pMag
                     /note="rapidly heterodimerizes with nMagHigh1 upon blue 
                     light irradiation to form a complex with slow switch-off 
                     kinetics"
                     /translation="HTLYAPGGYDIMGYLIQIMNRPNPQVELGPVDTSCALILCDLKQK
                     DTPIVYASEAFLYMTGYSNAEVLGRNCRFLQSPDGMVKPKSTRKYVDSNTINTMRKAID
                     RNAEVQVEVVNFKKNGQRFVNFLTMIPVRDETGEYRYSMGFQCETE"
     polyA_signal    3655..3776
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3692..3711)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3746..3765
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3783..4238)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3920..3941)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4132..4151
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4265..4369
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4371..4728
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4371..4562
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4394..4414)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4579..4714
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4763..5557
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4763..5557
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4817..4836)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5427..5446
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5745..5764)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5789..5836
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6165..6753
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6654..6673
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcacca
      541 tggctgctgc caagagagtt aagctggacg aggggggtgg tggatcaatg gtgagcaagg
      601 gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg
      661 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
      721 gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca
      781 tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc
      841 ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
      901 aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct
      961 acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga
     1021 ccatgggctg ggaggcctcc tccgagcgga tgtactccgg atactgtctg gatctgaaaa
     1081 cccaggttca gacaccgcag ggtatgaaag aaatttcaaa tattcaggtg ggtgatctgg
     1141 ttctgagcaa tactggttat aatgaagtgc tgaatgtgtt tccgaaaagc aaaaaaaaaa
     1201 gctataaaat caccctggaa gatggcaaag aaatcatttg tagcgaagaa cacctgtttc
     1261 cgacccagac tggtgaaatg aatattagcg gtggtctgaa agaaggtatg tgcctgtatg
     1321 ttaaaaagct tggtggagga ggttctaaca ttgatgagtt ggccctcaaa ttcgcaggac
     1381 tcgacctggg aggtggtgga tcaggaggag gcggttctat ggtgagcaag ggcgaggagc
     1441 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     1501 tcagcgtgcg cggcgagggc gagggcgatg ccaccaacgg caagctgacc ctgaagttca
     1561 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     1621 gcgtgcagtg cttcagccgc taccccgacc acatgaagcg ccacgacttc ttcaagtccg
     1681 ccatgcccga aggctacgtc caggagcgca ccatcagctt caaggacgac ggcacctaca
     1741 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     1801 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aacttcaaca
     1861 gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga
     1921 tccgccacaa cgtggaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     1981 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccaagc
     2041 tgagcaaaga tcccaacgag aaacgcgatc acatggtcct gctggagttc gtgaccgccg
     2101 ccgggatcac tctcggcatg gacgagctgt acaagggagg tggtggatca ggaggaggtg
     2161 gttctaccgg tgaaatgatg ctgaaaaaaa tcctgaaaat cgaggaactg gatgaacgcg
     2221 gtaccggcag cggaggtcat accttgtatg cgccgggggg ttatgacatc atgggttacc
     2281 tcatacagat catgaatagg ccgaacccac aagtggaact cggacccgtc gatacctcct
     2341 gcgctctcat tctgtgtgac cttaagcaga aggatacccc tatcgtgtac gcctccgagg
     2401 catttctgta catgacaggg tactcgaacg ccgaagtgct gggacggaac tgccgcttcc
     2461 tgcaaagccc ggatggaatg gtgaagccta agtcaacccg caaatacgtg gactccaaca
     2521 ctatcaacac catgcgcaag gccattgacc gcaatgctga ggtgcaagtg gaagtggtga
     2581 acttcaagaa gaatggacag cgcttcgtca acttcctgac tatgatcccc gtgcgcgacg
     2641 agaccggcga atatcggtac agcatggggt ttcagtgtga gacagagggc ggctcaggag
     2701 gcagcggcgg ttctggaggt tccggtggcg gctcaggagg tagcggaggg tctcacactc
     2761 tttacgcccc tggaggatac gacattatgg gatatttgat tcagattatg aaccgcccaa
     2821 accctcaggt cgaactgggg cctgtggaca cgtcatgtgc cctgatcctg tgcgatctga
     2881 agcaaaagga cactccgatt gtctacgcct cggaagcctt cttgtatatg accggataca
     2941 gcaatgcaga ggtgctcggt aggaactgca gattcctgca gtcccccgac gggatggtga
     3001 aaccaaagtc cactcgcaaa tatgtggact cgaacacgat caatacaatg cggaaggcca
     3061 tcgaccggaa cgccgaggtc caggtggagg tggtcaactt taagaagaac ggccagcggt
     3121 tcgtgaactt tctcaccatg attccggtcc gggatgaaac cggagagtac agatactcca
     3181 tgggattcca gtgcgaaacc gaagggtcag gatccgaact gattgatatt gaagttagcg
     3241 gtaaccacct gttctatgcc aatgatattc tgacccataa tagctcgagc cccgaggacg
     3301 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     3361 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     3421 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     3481 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag taagcggccg
     3541 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     3601 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     3661 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3721 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
     3781 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     3841 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     3901 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     3961 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     4021 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     4081 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     4141 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     4201 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     4261 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4321 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     4381 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4441 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4501 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4561 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4621 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     4681 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     4741 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4801 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4861 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4921 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     4981 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     5041 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     5101 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     5161 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     5221 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     5281 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     5341 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     5401 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     5461 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5521 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     5581 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5641 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5701 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     5761 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     5821 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     5881 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     5941 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     6001 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6061 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6121 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6181 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6241 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6301 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6361 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6421 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6481 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6541 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6601 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6661 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6721 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6781 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6841 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt
     6901 agttcatagc ccatatatgg agttccg
//
