LOCUS       sequence-417727-        5416 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NIR light-activated intein-based gene expression system (C-part).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5416)
  AUTHORS   Karasev MM, Verkhusha VV, Shcherbakova DM
  TITLE     Near-Infrared Optogenetic Module for Conditional Protein Splicing.
  JOURNAL   J Mol Biol. 2023 Dec 15;435(24):168360. doi: 
            10.1016/j.jmb.2023.168360. Epub 2023 Nov 8.
  PUBMED    37949312
REFERENCE   2  (bases 1 to 5416)
  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: "J Mol 
            Biol."; date: "2023-12-15"; volume: "435(24):168360. doi"; pages: " 
            10.1016/j.jmb.2023.168360. Epub 2023 Nov 8"
COMMENT     Sequence Label: ACE16-NLS
FEATURES             Location/Qualifiers
     source          1..5416
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        764..1067
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1068..1271
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1222..1242
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1699..1932
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     CDS             1933..1959
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2011..2064
                     /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(2065..2067,2068..2070,2071..2784)
                     /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(2110..2131)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2371..2390)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2718..2739
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2907..3028
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2944..2963)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2998..3017
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3035..3490)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3172..3193)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3384..3403
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3517..3621
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3623..3980
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3623..3814
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3646..3666)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3831..3966
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4015..4809
                     /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             4015..4809
                     /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(4069..4088)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4679..4698
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4997..5016)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5041..5088
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
      721 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      781 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      841 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      901 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      961 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1021 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1081 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     1141 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     1201 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     1261 taagcagagc tggtttagtg aaccgtcaga tccgctagta gatctagcac catgaagttt
     1321 aacaaggaga agtggcaggc tattaaggag attgccttcc tgcctaacct gaaccagccc
     1381 cagcacctgg cttttattgt gtctctgctg gacgaccctt ctcagtctgc taacctgctg
     1441 gctgaggcta agaagctgaa cgacgctcag gctcctaagg gatccgaact gattgatatt
     1501 gaagttagcg gtaaccacct gttctatgcc aatgatattc tgacccataa tagctcgagc
     1561 gcgcgtacga aaaacaatta cgggtctacc atcgagggcc tgctcgatct cccggacgac
     1621 gacgcccccg aagaggcggg gctggcggct ccgcgcctgt cctttctccc cgcgggacac
     1681 acgcgcagac tgtcgacggc ccccccgacc gatgtcagcc tgggggacga gctccactta
     1741 gacggcgagg acgtggcgat ggcgcatgcc gacgcgctag acgatttcga tctggacatg
     1801 ttgggggacg gggattcccc gggtccggga tttacccccc acgactccgc cccctacggc
     1861 gctctggata tggccgactt cgagtttgag cagatgttta ccgatgccct tggaattgac
     1921 gagtacggtg ggtaccctta cgatgtgccc gattacgccg gatctgctgc tgccaagaga
     1981 gttaagctgg acgaggaatt cggtagcggc gagggcagag gaagtctgct aacatgcggt
     2041 gatgtcgagg agaatcctgg cccaatggtg agcaagggcg aggagctgtt caccggggtg
     2101 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     2161 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     2221 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     2281 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     2341 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     2401 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     2461 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     2521 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     2581 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     2641 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaagctgag caaagacccc
     2701 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     2761 ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca
     2821 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     2881 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     2941 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3001 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     3061 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     3121 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     3181 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     3241 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     3301 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     3361 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     3421 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     3481 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3541 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3601 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     3661 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3721 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3781 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3841 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3901 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3961 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4021 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4081 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4141 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     4201 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4261 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4321 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4381 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4441 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4501 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     4561 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4621 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4681 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4741 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4801 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4861 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4921 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4981 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     5041 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     5101 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     5161 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     5221 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     5281 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5341 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5401 agatcaaagg atcttc
//
