LOCUS       sequence-417725-        7317 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NIR light-activated intein-based gene expression system (N-part) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7317)
  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 7317)
  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: TND-NES
FEATURES             Location/Qualifiers
     source          1..7317
                     /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             1306..1347
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1348..1965
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="SRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHVK
                     NKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTRP
                     TEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTTD
                     SMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(1418..1437)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     polyA_signal    4808..4929
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4845..4864)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4899..4918
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4936..5391)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5073..5094)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5285..5304
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5418..5522
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5524..5881
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5524..5715
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5547..5567)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5732..5867
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5916..6710
                     /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             5916..6710
                     /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(5970..5989)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6580..6599
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6898..6917)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6942..6989
                     /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 tccgctagca ccatgggcaa gcccattccc
     1321 aatcccttgc tggggttgga ctccacctcc agattagata aaagtaaagt gattaacagc
     1381 gcattagagc tgcttaatga ggtcggaatc gaaggtttaa caacccgtaa actcgcccag
     1441 aagctaggtg tagagcagcc tacattgtat tggcatgtaa aaaataagcg ggctttgctc
     1501 gacgccttag ccattgagat gttagatagg caccatactc acttttgccc tttagaaggg
     1561 gaaagctggc aagatttttt acgtaataac gctaaaagtt ttagatgtgc tttactaagt
     1621 catcgcgatg gagcaaaagt acatttaggt acacggccta cagaaaaaca gtatgaaact
     1681 ctcgaaaatc aattagcctt tttatgccaa caaggttttt cactagagaa tgcattatat
     1741 gcactcagcg ctgtggggca ttttacttta ggttgcgtat tggaagatca agagcatcaa
     1801 gtcgctaaag aagaaaggga aacacctact actgatagta tgccgccatt attacgacaa
     1861 gctatcgaat tattcgatca ccaaggtgca gagccagcct tcttattcgg ccttgaattg
     1921 attatatgcg gattagaaaa acaacttaaa tgtgaaagtg ggtccgcgta cagccgctcc
     1981 ggatactgtc tggatctgaa aacccaggtt cagacaccgc agggtatgaa agaaatttca
     2041 aatattcagg tgggtgatct ggttctgagc aatactggtt ataatgaagt gctgaatgtg
     2101 tttccgaaaa gcaaaaaaaa aagctataaa atcaccctgg aagatggcaa agaaatcatt
     2161 tgtagcgaag aacacctgtt tccgacccag actggtgaaa tgaatattag cggtggtctg
     2221 aaagaaggta tgtgcctgta tgttaaagaa atgatgctga aaaaaatcct gaaaatcgag
     2281 gaactggatg aacgcggtac cagtgctggt gggtccgccg gcgggagtgc tggcggtagt
     2341 gccggcggta gtctcgaggg ctccatgagc cgcgaccccc tgccattctt cccccccctg
     2401 tacctgggag gacccgagat taccaccgaa aactgtgaac gagagcctat ccacatcccc
     2461 ggaagtattc agcctcatgg cgccctgctg actgctgatg gccacagtgg ggaagtgctg
     2521 cagatgtcac tgaacgccgc taccttcctg gggcaggagc ctactgtgct gaggggacag
     2581 accctggcag cactgctgcc tgaacagtgg ccagcactgc aggctgcact gccacctggc
     2641 tgccccgatg ccctgcagta cagggctact ctggactggc ctgcagctgg acacctgagc
     2701 ctgaccgtcc atcgcgtggg ggaactgctg atcctggagt tcgaaccaac agaggcttgg
     2761 gatagcactg gcccccacgc actgcggaac gctatgtttg cactggagtc cgcccccaat
     2821 ctgagagcac tggccgaagt cgccactcag accgtgcggg agctgacagg gttcgataga
     2881 gtcatgctgt acaagtttgc ccctgacgct actggagaag tgatcgctga agcacggaga
     2941 gaggggctgc acgctttcct gggacatcga tttccagcct ccgacattcc agcacaggct
     3001 cgggcactgt ataccaggca cctgctgcga ctgacagcag atactcgcgc agcagctgtc
     3061 ccactggacc ccgtgctgaa ccctcagacc aatgcaccaa cacctctggg aggagcagtg
     3121 ctgagagcaa cctctccaat gcatatgcag tatctgagga atatgggagt cggcagctcc
     3181 ctgtctgtga gtgtggtcgt gggaggacag ctgtggggac tgattgcctg ccaccatcag
     3241 acaccctacg tgctgccacc cgatctgagg accacactgg agtatctggg gcgcctgctg
     3301 tctctgcagg tccaggtgaa agaagcagcc gacgtcgctg cattccgaca gagtctgcga
     3361 gagcaccatg cacgggtggc cctggccgct gcacactcac tgagccctca tgacacactg
     3421 agcgatccag ctctggacct gctgggactg atgagagcag gaggactgat cctgagattt
     3481 gaaggcaggt ggcagactct gggagaggtc cctccagcac cagctgtgga cgctctgctg
     3541 gcatggctgg agacccagcc aggagctctg gtgcagacag atgcactggg acagctgtgg
     3601 ccagcaggag cagacctggc tccttcagca gctggactgc tggccatttc cgtcggagaa
     3661 ggatggtctg agtgtctggt gtggctgcgc ccagaactgc gactggaggt ggcttgggga
     3721 ggagcaaccc ctgatcaggc aaaggacgat ctgggaccac gccacagctt tgacacatac
     3781 ctggaggaaa aacgaggcta tgccgagccc tggcacccag gggaaattga ggaggcacag
     3841 gatctgagag acacactgac aggggcactg ggggagagac tgtccgtgat tcgcgatctc
     3901 aaccgggctc tgacccagtc caatgccgag tggcgtcagt atggcttcgt catcagtcac
     3961 cacatgcagg aacccgtcag actgatctcc cagtttgccg agcttctgac tcgccaaccc
     4021 agagcccagg atggcagccc ggactctccc cagacagagc gtatcaccgg ctttctgctc
     4081 agggaaacct cacgactccg ctctctgacc caggacttgc acacgtacac tgcgctgctg
     4141 tcagcacctc ctccagttag gaggccaaca ccacttgggc gagttgtgga cgatgtgctg
     4201 caggacctcg aaccacggat tgccgataca ggggcctcca tagaggtagc tcccgaactg
     4261 cccgtcattg ccgctgatgc tggcctcttg cgcgaccttc tgctgcatct gattggtaac
     4321 gccttgacct tcggaggacc agagcctcgg attgctgtga gaaccgaacg gcaaggtgcc
     4381 ggctggagca tcgctgtgag cgatcagggt gcggggatag cgccggagta tcaggagagg
     4441 atcttcttgc tcttccagag actcggctct ctggacgagg cactgggcaa tggacttggg
     4501 cttcccctgt gtcggaagat cgccgaactg catggtggca cactgactgt ggagagtgca
     4561 cctggcgaag gaagcacgtt taggtgctgg ctgcctgatg ccggaccact gcctggagca
     4621 gcagacgcta ctagtaattc taacgaactc gctttgaaat tggcgggctt ggacatcaac
     4681 aagtaagcgg ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     4741 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     4801 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4861 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4921 tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     4981 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     5041 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     5101 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     5161 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     5221 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     5281 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     5341 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     5401 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     5461 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     5521 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     5581 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     5641 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     5701 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     5761 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     5821 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     5881 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     5941 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     6001 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     6061 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     6121 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     6181 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     6241 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     6301 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     6361 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     6421 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     6481 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     6541 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     6601 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     6661 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     6721 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     6781 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     6841 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     6901 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     6961 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     7021 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     7081 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     7141 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     7201 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     7261 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttc
//
