LOCUS       sequence-481810-        6820 bp    DNA     circular SYN 08-AUG-2025
DEFINITION  Blue_light-induced reconstitution of split mCherry..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6820)
  TITLE     Photoswitchable intein for light control of covalent protein binding
            and cleavage
REFERENCE   2  (bases 1 to 6820)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 8, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: PS Intein
FEATURES             Location/Qualifiers
     source          1..6820
                     /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(685..703)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     953..972
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1860..2309
                     /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"
     CDS             2652..2705
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(2706..2708,2709..2711,2712..3425)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2751..2772)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3012..3031)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3359..3380
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3548..3669
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3585..3604)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3639..3658
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3676..4131)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3813..3834)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4025..4044
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4158..4262
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4264..4621
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4264..4455
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4287..4307)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4472..4607
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4656..5450
                     /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             4656..5450
                     /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(4710..4729)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5320..5339
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5638..5657)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5682..5729
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6058..6646
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6547..6566
                     /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 tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg
      601 tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc
      661 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct
      721 tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc
      781 ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg
      841 tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg
      901 gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa
      961 tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtactcc ggatactgtc
     1021 tggatctgaa aacccaggtt cagacaccgc agggtatgaa agaaatttca aatattcagg
     1081 tgggtgatct ggttctgagc aatactggtt ataatgaagt gctgaatgtg tttccgaaaa
     1141 gcaaaaaaaa aagctataaa atcaccctgg aagatggcaa agaaatcatt tgtagcgaag
     1201 aacacctgtt tccgacccag actggtgaaa tgaatattag cggtggtctg aaagaaggta
     1261 tgtgcctgta tgttaaagaa atgatgctga aaaaaatcct gaaaatcgag gaactggatg
     1321 aacgcggtac cggcagcgga ggtcatacct tgtatgcgcc ggggggttat gacatcatgg
     1381 gttacctcat acagatcatg aataggccga acccacaagt ggaactcgga cccgtcgata
     1441 cctcctgcgc tctcattctg tgtgacctta agcagaagga tacccctatc gtgtacgcct
     1501 ccgaggcatt tctgtacatg acagggtact cgaacgccga agtgctggga cggaactgcc
     1561 gcttcctgca aagcccggat ggaatggtga agcctaagtc aacccgcaaa tacgtggact
     1621 ccaacactat caacaccatg cgcaaggcca ttgaccgcaa tgctgaggtg caagtggaag
     1681 tggtgaactt caagaagaat ggacagcgct tcgtcaactt cctgactatg atccccgtgc
     1741 gcgacgagac cggcgaatat cggtacagca tggggtttca gtgtgagaca gagggcggct
     1801 caggaggcag cggcggttct ggaggttccg gtggcggctc aggaggtagc ggagggtctc
     1861 acactcttta cgcccctgga ggatacgaca ttatgggata tttgattcag attatgaacc
     1921 gcccaaaccc tcaggtcgaa ctggggcctg tggacacgtc atgtgccctg atcctgtgcg
     1981 atctgaagca aaaggacact ccgattgtct acgcctcgga agccttcttg tatatgaccg
     2041 gatacagcaa tgcagaggtg ctcggtagga actgcagatt cctgcagtcc cccgacggga
     2101 tggtgaaacc aaagtccact cgcaaatatg tggactcgaa cacgatcaat acaatgcgga
     2161 aggccatcga ccggaacgcc gaggtccagg tggaggtggt caactttaag aagaacggcc
     2221 agcggttcgt gaactttctc accatgattc cggtccggga tgaaaccgga gagtacagat
     2281 actccatggg attccagtgc gaaaccgaag ggtcaggatc cgaactgatt gatattgaag
     2341 ttagcggtaa ccacctgttc tatgccaatg atattctgac ccataatagc tcgagccccg
     2401 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact
     2461 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     2521 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
     2581 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaaggaat
     2641 tcggtagcgg cgagggcaga ggaagtctgc taacatgcgg tgatgtcgag gagaatcctg
     2701 gcccaatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     2761 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     2821 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     2881 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     2941 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     3001 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     3061 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     3121 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     3181 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     3241 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     3301 accactacct gagcacccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca
     3361 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     3421 agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     3481 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     3541 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3601 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3661 tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     3721 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     3781 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     3841 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     3901 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     3961 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     4021 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     4081 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     4141 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4201 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4261 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     4321 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     4381 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4441 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4501 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4561 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4621 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4681 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4741 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4801 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     4861 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4921 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4981 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     5041 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     5101 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     5161 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     5221 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5281 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5341 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5401 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     5461 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5521 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5581 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     5641 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     5701 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     5761 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     5821 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     5881 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     5941 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6001 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6061 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6121 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6181 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6241 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6301 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6361 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6421 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6481 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6541 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6601 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6661 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6721 atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa
     6781 ttacggggtc attagttcat agcccatata tggagttccg
//
