LOCUS       sequence-417728-        7056 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NIR optogenetic module for conditional protein splicing of split 
            mCherry (N-part).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7056)
  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 7056)
  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: pN1-CMV-mCherryN-IntN-DrBphP (N23)
FEATURES             Location/Qualifiers
     source          1..7056
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(188..207)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    232..279
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      608..1196
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1097..1116
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1371..1674
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1675..1878
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1829..1849
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(2055..2073)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2323..2342
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5154..5275
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5191..5210)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5245..5264
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5282..5737)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5419..5440)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5631..5650
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5764..5868
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5870..6227
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5870..6061
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5893..5913)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6078..6213
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6262..7056
                     /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             6262..7056
                     /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(6316..6335)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6926..6945
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
       61 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
      121 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
      181 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
      241 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
      301 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
      361 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
      421 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
      481 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
      541 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
      601 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
      661 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
      721 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
      781 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
      841 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
      901 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
      961 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     1021 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     1081 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     1141 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     1201 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     1261 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa
     1321 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     1381 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     1441 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     1501 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     1561 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     1621 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     1681 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     1741 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     1801 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     1861 gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcacca tggtgagcaa
     1921 gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga
     1981 gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga
     2041 gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga
     2101 catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat
     2161 ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt
     2221 cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat
     2281 ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa
     2341 gaccatgggc tgggaggcct cctccgagcg gatgtactcc ggatactgtc tggatctgaa
     2401 aacccaggtt cagacaccgc agggtatgaa agaaatttca aatattcagg tgggtgatct
     2461 ggttctgagc aatactggtt ataatgaagt gctgaatgtg tttccgaaaa gcaaaaaaaa
     2521 aagctataaa atcaccctgg aagatggcaa agaaatcatt tgtagcgaag aacacctgtt
     2581 tccgacccag actggtgaaa tgaatattag cggtggtctg aaagaaggta tgtgcctgta
     2641 tgttaaagaa atgatgctga aaaaaatcct gaaaatcgag gaactggatg aacgcggtac
     2701 cagtgctggt gggtccgccg gcgggagtgc tggcggtagt gccggcggta gtctcgaggg
     2761 ctccatgagc cgcgaccccc tgccattctt cccccccctg tacctgggag gacccgagat
     2821 taccaccgaa aactgtgaac gagagcctat ccacatcccc ggaagtattc agcctcatgg
     2881 cgccctgctg actgctgatg gccacagtgg ggaagtgctg cagatgtcac tgaacgccgc
     2941 taccttcctg gggcaggagc ctactgtgct gaggggacag accctggcag cactgctgcc
     3001 tgaacagtgg ccagcactgc aggctgcact gccacctggc tgccccgatg ccctgcagta
     3061 cagggctact ctggactggc ctgcagctgg acacctgagc ctgaccgtcc atcgcgtggg
     3121 ggaactgctg atcctggagt tcgaaccaac agaggcttgg gatagcactg gcccccacgc
     3181 actgcggaac gctatgtttg cactggagtc cgcccccaat ctgagagcac tggccgaagt
     3241 cgccactcag accgtgcggg agctgacagg gttcgataga gtcatgctgt acaagtttgc
     3301 ccctgacgct actggagaag tgatcgctga agcacggaga gaggggctgc acgctttcct
     3361 gggacatcga tttccagcct ccgacattcc agcacaggct cgggcactgt ataccaggca
     3421 cctgctgcga ctgacagcag atactcgcgc agcagctgtc ccactggacc ccgtgctgaa
     3481 ccctcagacc aatgcaccaa cacctctggg aggagcagtg ctgagagcaa cctctccaat
     3541 gcatatgcag tatctgagga atatgggagt cggcagctcc ctgtctgtga gtgtggtcgt
     3601 gggaggacag ctgtggggac tgattgcctg ccaccatcag acaccctacg tgctgccacc
     3661 cgatctgagg accacactgg agtatctggg gcgcctgctg tctctgcagg tccaggtgaa
     3721 agaagcagcc gacgtcgctg cattccgaca gagtctgcga gagcaccatg cacgggtggc
     3781 cctggccgct gcacactcac tgagccctca tgacacactg agcgatccag ctctggacct
     3841 gctgggactg atgagagcag gaggactgat cctgagattt gaaggcaggt ggcagactct
     3901 gggagaggtc cctccagcac cagctgtgga cgctctgctg gcatggctgg agacccagcc
     3961 aggagctctg gtgcagacag atgcactggg acagctgtgg ccagcaggag cagacctggc
     4021 tccttcagca gctggactgc tggccatttc cgtcggagaa ggatggtctg agtgtctggt
     4081 gtggctgcgc ccagaactgc gactggaggt ggcttgggga ggagcaaccc ctgatcaggc
     4141 aaaggacgat ctgggaccac gccacagctt tgacacatac ctggaggaaa aacgaggcta
     4201 tgccgagccc tggcacccag gggaaattga ggaggcacag gatctgagag acacactgac
     4261 aggggcactg ggggagagac tgtccgtgat tcgcgatctc aaccgggctc tgacccagtc
     4321 caatgccgag tggcgtcagt atggcttcgt catcagtcac cacatgcagg aacccgtcag
     4381 actgatctcc cagtttgccg agcttctgac tcgccaaccc agagcccagg atggcagccc
     4441 ggactctccc cagacagagc gtatcaccgg ctttctgctc agggaaacct cacgactccg
     4501 ctctctgacc caggacttgc acacgtacac tgcgctgctg tcagcacctc ctccagttag
     4561 gaggccaaca ccacttgggc gagttgtgga cgatgtgctg caggacctcg aaccacggat
     4621 tgccgataca ggggcctcca tagaggtagc tcccgaactg cccgtcattg ccgctgatgc
     4681 tggcctcttg cgcgaccttc tgctgcatct gattggtaac gccttgacct tcggaggacc
     4741 agagcctcgg attgctgtga gaaccgaacg gcaaggtgcc ggctggagca tcgctgtgag
     4801 cgatcagggt gcggggatag cgccggagta tcaggagagg atcttcttgc tcttccagag
     4861 actcggctct ctggacgagg cactgggcaa tggacttggg cttcccctgt gtcggaagat
     4921 cgccgaactg catggtggca cactgactgt ggagagtgca cctggcgaag gaagcacgtt
     4981 taggtgctgg ctgcctgatg ccggaccact gcctggagca gcagacgctt aagcggccgc
     5041 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     5101 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     5161 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5221 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg
     5281 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     5341 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     5401 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     5461 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     5521 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     5581 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     5641 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     5701 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     5761 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     5821 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     5881 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     5941 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     6001 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     6061 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     6121 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     6181 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     6241 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     6301 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     6361 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     6421 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     6481 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     6541 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     6601 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     6661 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     6721 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     6781 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     6841 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     6901 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     6961 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     7021 atcgccttct atcgccttct tgacgagttc ttctga
//
