LOCUS       sequence-334945-        6581 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NLS-Gal4(1-65)-iLight(human codon optimized)-VP16-T2A-mTagBFP2 
            construct expression in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6581)
  AUTHORS   Kaberniuk AA, Baloban M, Monakhov MV, Shcherbakova DM, Verkhusha VV
  TITLE     Single-component near-infrared optogenetic systems for gene 
            transcription regulation.
  JOURNAL   Nat Commun. 2021 Jun 23;12(1):3859. doi: 10.1038/s41467-021-24212-7.
  PUBMED    34162879
REFERENCE   2  (bases 1 to 6581)
  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: "Nat 
            Commun."; date: "2021-06-23"; volume: "12(1):3859. doi"; pages: " 
            10.1038/s41467-021-24212-7"
COMMENT     Sequence Label: pCMVd1-NLS-GAL4-iLight-VP16-T2A-mTagBFP2
FEATURES             Location/Qualifiers
     source          1..6581
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..100
                     /label=CMVd1 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields slightly reduced expression 
                     relative to the full-length promoter"
     primer_bind     51..71
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2268..2501
                     /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             2520..2573
                     /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             2580..3278
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     polyA_signal    3413..3534
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3450..3469)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3504..3523
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3541..3996)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3678..3699)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3890..3909
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4023..4127
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4129..4486
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4129..4320
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4152..4172)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4337..4472
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4521..5315
                     /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             4521..5315
                     /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(4575..4594)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5185..5204
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5503..5522)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5547..5594
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5923..6511
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6412..6431
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
       61 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
      121 ccgctagcat ggcgcgcaag aagaagagga agtccggagg tggtggaaag ctactgtctt
      181 ctatcgaaca agcatgcgat atttgccgac ttaaaaagct caagtgctcc aaagaaaaac
      241 cgaagtgcgc caagtgtctg aagaacaact gggagtgtcg ctactctccc aaaaccaaaa
      301 ggtctccgct gactagggca catctgacag aagtggaatc aaggctagaa agactggaac
      361 agactagtgc tggtgggtcc gccggcggga gtgctggcgg tagtgccggc ggtgaattcg
      421 ccgccgacct gggctctgac gatatcagca agctgatcgc cgcctgcgat caggagccaa
      481 tccacatccc caatgccatc cagccatttg gcgccatgct gatcgtggag aaggacacac
      541 agcagatcgt gtacgcctct gccaacagcg ccgagtactt cagcgtggcc gacaatacca
      601 tccacgagct gtccgatttc aagcaggcca acatcaattc tctgctgccc gagcagctga
      661 tcagcggcct gacatccgcc atctctgaga acgagcctat ctgggtggag accgacaggc
      721 tgagctttct gggctggcgc cacgagaact actatatcat cgaggtggag agataccacg
      781 tgcagacatc caattggttc gagatccagt ttcagcgggc cttccagaag ctgagaaact
      841 gtaagaccca caacgatctg atcaataccc tgacacggct gatccaggag atcagcggct
      901 acgacagagt gatgatctat cagttcgatc ccgagtggaa tggcagagtg atcgccgaga
      961 gcgtgagaca gctgtttacc tccatgctga accaccactt cccagcctct gacatccctg
     1021 cacaggccag ggccatgtac agcatcaacc caatccgcat catccccgat gtgaatgccg
     1081 agccccagcc tctgcacatg atccacaagc cacagaacac agaggccgtg aatctgtcct
     1141 gcggcgtgct gagggccgtg tctccactgc acatgcagta tctgcgcaac tttggcgtgt
     1201 ctgccagcac ctccatcggc atcttcaatg aggacaagct gtggggcatc gtggcctgtc
     1261 accacacaaa gcctagggcc atcggccgga gaatcaggca cctgctggtg cgcaccgtgg
     1321 agtttgcagc agagcgcctg tggctgatcc actccaggaa tgtggagcgg tacatggtga
     1381 cagtgcaggc agcccgggag cagctgtcta ccacagccga cgataagcac agctcccacg
     1441 agatcgtgat cgagcacgcc gccgactggt gcaagctgtt ccggtgtgat ggcgtgggct
     1501 acctgagagg cgaggagctg accacatatg gcgagacccc tgatcagacc acaatcaaca
     1561 agctggtgga gtggctggag gagaatggca agaagagcct gttttggcac tcccacatgc
     1621 tgaaggagga cgcacctgga ctgctgccag atggcagccg gttcgcagga ctgctggcca
     1681 tcccactgaa gtctgacgcc gatctgttta gctacctgct gctgttcagg gtggcacaga
     1741 acgaggtgcg cacatgggca ggcaagcctg agaagctgtc cgtggagacc tctacaggca
     1801 ccatggtggg cccacggaag tcttttgagg cctggcagga cgaggtgagc ggcaagtccc
     1861 agccttggag aaccgcacag ctgtatgcag cccgggacat cgcccgggac ctgctgatcg
     1921 tggccgatag catgcagctg aacctgctga atgaccagct ggccgatgcc aacgagaatc
     1981 tggagaagct ggcctccttc gacgatctga cctctggcgg cggtaccagc gggggtggtg
     2041 gatcaggtgg aggaggttct ggaggtggtg gatcaggagg aggtggttct ggaggtggtg
     2101 gatcaggagg aagatctgcg tacagccgcg cgcgtacgaa aaacaattac gggtctacca
     2161 tcgagggcct gctcgatctc ccggacgacg acgcccccga agaggcgggg ctggcggctc
     2221 cgcgcctgtc ctttctcccc gcgggacaca cgcgcagact gtcgacggcc cccccgaccg
     2281 atgtcagcct gggggacgag ctccacttag acggcgagga cgtggcgatg gcgcatgccg
     2341 acgcgctaga cgatttcgat ctggacatgt tgggggacgg ggattcccct ggtccgggat
     2401 ttacccccca cgactccgcc ccctacggcg ctctggatat ggccgacttc gagtttgagc
     2461 agatgtttac cgatgccctt ggaattgacg agtacggtgg ggctcgagat aggcgcgccg
     2521 agggcagagg aagtctgcta acatgcggtg acgtcgagga gaatcctggc ccaactagta
     2581 gcgagctgat taaggagaac atgcacatga agctgtacat ggagggcacc gtggacaacc
     2641 atcacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc cagaccatga
     2701 gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg gctactagct
     2761 tcctctacgg cagcaagacc ttcatcaacc acacccaggg catccccgac ttcttcaagc
     2821 agtccttccc tgagggcttc acatgggaga gagtcaccac atacgaagac gggggcgtgc
     2881 tgaccgctac ccaggacacc agcctccagg acggctgcct catctacaac gtcaagatca
     2941 gaggggtgaa cttcacatcc aacggccctg tgatgcagaa gaaaacactc ggctgggagg
     3001 ccttcaccga gacgctgtac cccgctgacg gcggcctgga aggcagaaac gacatggccc
     3061 tgaagctcgt gggcgggagc catctgatcg caaacgccaa gaccacatat agatccaaga
     3121 aacccgctaa gaacctcaag atgcctggcg tctactatgt ggactacaga ctggaaagaa
     3181 tcaaggaggc caacaacgag acctacgtcg agcagcacga ggtggcagtg gccagatact
     3241 gcgacctccc tagcaaactg gggcacaagc ttaattaacc cgggatccac cggggccgcg
     3301 actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     3361 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     3421 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3481 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt
     3541 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     3601 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     3661 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     3721 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     3781 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     3841 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     3901 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     3961 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     4021 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4081 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     4141 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     4201 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4261 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4321 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4381 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     4441 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     4501 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4561 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4621 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4681 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     4741 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4801 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4861 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4921 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4981 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     5041 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5101 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5161 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5221 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5281 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5341 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5401 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5461 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     5521 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     5581 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     5641 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     5701 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     5761 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5821 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5881 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5941 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6001 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6061 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6121 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6181 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6241 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6301 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6361 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6421 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6481 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6541 cctggccttt tgctggcctt ttgctcacat gttctttcct g
//
