LOCUS       sequence-481813-        6886 bp    DNA     circular SYN 08-AUG-2025
DEFINITION  Blue-light-induced reconstitution of split mCherry. A T2A peptide is
            inserted at the natural split site of gp41-1..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6886)
  TITLE     Photoswitchable intein for light control of covalent protein binding
            and cleavage
REFERENCE   2  (bases 1 to 6886)
  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: Split PS Intein
FEATURES             Location/Qualifiers
     source          1..6886
                     /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             1287..1340
                     /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             1926..2375
                     /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             2718..2771
                     /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(2772..2774,2775..2777,2778..3491)
                     /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(2817..2838)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3078..3097)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3425..3446
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3614..3735
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3651..3670)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3705..3724
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3742..4197)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3879..3900)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4091..4110
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4224..4328
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4330..4687
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4330..4521
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4353..4373)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4538..4673
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4722..5516
                     /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             4722..5516
                     /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(4776..4795)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5386..5405
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5704..5723)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5748..5795
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6124..6712
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6613..6632
                     /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 tgttaaaggc agcggcgagg gcaggggcag cctgctgacc tgcggcgacg
     1321 tggaggagaa ccccggcccc ggcgaaatga tgctgaaaaa aatcctgaaa atcgaggaac
     1381 tggatgaacg cggtaccggc agcggaggtc ataccttgta tgcgccgggg ggttatgaca
     1441 tcatgggtta cctcatacag atcatgaata ggccgaaccc acaagtggaa ctcggacccg
     1501 tcgatacctc ctgcgctctc attctgtgtg accttaagca gaaggatacc cctatcgtgt
     1561 acgcctccga ggcatttctg tacatgacag ggtactcgaa cgccgaagtg ctgggacgga
     1621 actgccgctt cctgcaaagc ccggatggaa tggtgaagcc taagtcaacc cgcaaatacg
     1681 tggactccaa cactatcaac accatgcgca aggccattga ccgcaatgct gaggtgcaag
     1741 tggaagtggt gaacttcaag aagaatggac agcgcttcgt caacttcctg actatgatcc
     1801 ccgtgcgcga cgagaccggc gaatatcggt acagcatggg gtttcagtgt gagacagagg
     1861 gcggctcagg aggcagcggc ggttctggag gttccggtgg cggctcagga ggtagcggag
     1921 ggtctcacac tctttacgcc cctggaggat acgacattat gggatatttg attcagatta
     1981 tgaaccgccc aaaccctcag gtcgaactgg ggcctgtgga cacgtcatgt gccctgatcc
     2041 tgtgcgatct gaagcaaaag gacactccga ttgtctacgc ctcggaagcc ttcttgtata
     2101 tgaccggata cagcaatgca gaggtgctcg gtaggaactg cagattcctg cagtcccccg
     2161 acgggatggt gaaaccaaag tccactcgca aatatgtgga ctcgaacacg atcaatacaa
     2221 tgcggaaggc catcgaccgg aacgccgagg tccaggtgga ggtggtcaac tttaagaaga
     2281 acggccagcg gttcgtgaac tttctcacca tgattccggt ccgggatgaa accggagagt
     2341 acagatactc catgggattc cagtgcgaaa ccgaagggtc aggatccgaa ctgattgata
     2401 ttgaagttag cggtaaccac ctgttctatg ccaatgatat tctgacccat aatagctcga
     2461 gccccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
     2521 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
     2581 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg
     2641 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     2701 aggaattcgg tagcggcgag ggcagaggaa gtctgctaac atgcggtgat gtcgaggaga
     2761 atcctggccc aatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     2821 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     2881 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     2941 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     3001 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     3061 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     3121 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     3181 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     3241 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     3301 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     3361 ccgacaacca ctacctgagc acccagtcca agctgagcaa agaccccaac gagaagcgcg
     3421 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     3481 tgtacaagta aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt
     3541 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     3601 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     3661 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     3721 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     3781 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     3841 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     3901 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     3961 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     4021 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     4081 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     4141 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     4201 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4261 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4321 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     4381 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4441 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4501 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4561 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4621 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4681 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4741 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4801 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4861 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     4921 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4981 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     5041 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     5101 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     5161 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     5221 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     5281 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5341 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5401 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5461 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     5521 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5581 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5641 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     5701 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     5761 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     5821 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     5881 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     5941 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     6001 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6061 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6121 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6181 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6241 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     6301 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6361 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6421 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6481 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6541 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6601 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6661 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6721 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     6781 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt
     6841 aatcaattac ggggtcatta gttcatagcc catatatgga gttccg
//
