LOCUS       sequence-391271-        5666 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the synthetic photodegradable transcription factor 
            Gal4-degFusionRed-p65 in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5666)
  AUTHORS   Chernov KG, Manoilov KY, Oliinyk OS, Shcherbakova DM, Verkhusha VV
  TITLE     Photodegradable by Yellow-Orange Light degFusionRed Optogenetic 
            Module with Autocatalytically Formed Chromophore.
  JOURNAL   Int J Mol Sci. 2023 Mar 30;24(7):6526. doi: 10.3390/ijms24076526.
  PUBMED    37047499
REFERENCE   2  (bases 1 to 5666)
  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: "Int J Mol 
            Sci."; date: "2023-03-30"; volume: "24(7):6526. doi"; pages: " 
            10.3390/ijms24076526"
COMMENT     Sequence Label: pCMV-Gal4-degFusionRed-p65
FEATURES             Location/Qualifiers
     source          1..5666
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        48..239
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     196..216
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(241..345)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      372..827
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(459..478)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     669..690
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    834..955
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(845..864)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     899..918
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1076..1099)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(1100..1471)
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALLSQISS"
     promoter        complement(2841..3044)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2870..2890)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3045..3348
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3523..4111)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3603..3622)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4440..4487
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4512..4531
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4719..5513)
                     /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             complement(4719..5513)
                     /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(4830..4849)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5440..5459
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5548..5666,1..239))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5562..5666,1..31)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg
       61 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
      121 cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
      181 ctggggactt tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcagg
      241 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
      301 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      361 agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
      421 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      481 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      541 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      601 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      661 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      721 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      781 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttacg ccttaagata
      841 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
      901 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
      961 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     1021 caagtaaaac ctctacaaat gtggtatggc tgattatgat ctagagtgta cattacttgt
     1081 catcatcgtc tttgtagtcg gagctgatct gactcagcag ggctgagaag tccatgtccg
     1141 caatggagga gaagtcttca tctcctgaaa ggaggccatt ggggagcccc ggggccccca
     1201 gtggagcagg agctgggtcg gggggcctct gggcccctgt cactaggcga gttatagcct
     1261 cagggtactc catcagcatg ggctcagttg tgtggggggc cacaggtatg ccctggttca
     1321 gcagctgctg aaactcggag ttgtcgacgg atgccaggtc tgtgaacaca gctgggtctg
     1381 tgctgttgcc aagcaaggcc cccaggtctt catcatcaaa ctgcagctgc agcagggcct
     1441 ctgacagcgt tccttcccca gcctgggtgg gcttgggggc aggtggggcc acagcctgag
     1501 gagggcctgg ggctaggact gggacagggg ctggggcctg ggccagagct gataccatgg
     1561 ctggagcagg ggcaggggct ggagcctggg gcaggacttg gggaggggcc ggggccaagg
     1621 ccgaggcctg gctgatctgc ccagaaggaa acaccatggt gggaaactca tcatagttga
     1681 tggtgctcag ggatgacgta aagggatagg gctggggtgc tggcttgggg acagaagctg
     1741 agctgcggga aggcacagca atgcgtcgag gtggaggccg ggggtcggtg ggtccgctga
     1801 aaggactctt cttcatgatg ctcttgaagg tctcatatgt ccttttacgt ttctcctcaa
     1861 tccggtgacg atcgtctgta tctggcaggt actggaattc tttccctcca tcgcctgccc
     1921 cccctgttga gtatcttgcc actgccactt cgtgctgttc cacatatgtc tcatcgtcag
     1981 cttccttgat ccgctccagc cgatgatcga cattatacac gccaggcatc ttcaggttag
     2041 tggcgggctt cttgctccgg taggtagttc ccagattaca gatcaggtgt cccccgccca
     2101 cgtttttcag agccatccag caccttcccc tcagttctcc gtctgctgcg tacataagtt
     2161 cagttgaggg cacccacctc agggttttct tctgcatgac tgggccattg gcagggaagt
     2221 tcactcctct gaccttcacg ttatagatca ggcagccatc ctggagactt gtgtcctgtg
     2281 tggcggtcag gacgcctcca tcctcgtatg tggtcactcg ttcccaggtg aagccctcgg
     2341 ggaaagactg tttaaagaag tcggggatgc caggggggtg cttgataaaa gtgcgggacc
     2401 catacatgaa gctggtggcc agaatgtcaa atgcgaaggg cagtggcccg ccttcgacca
     2461 ctttgatcct catagtctgt gtcccctcgt aaggcttccc ttcgccctcg cttgtgcatt
     2521 tgaagtggtg gttgttgacg gttccctcca tatacagttt cataggcatg ttctccttaa
     2581 tcagctcgct catagatctg gtggcgatgg atctttccag tctttctagc cttgattcca
     2641 cttctgtcag atgtgcccta gtcagcggag accttttggt tttgggagag tagcgacact
     2701 cccagttgtt cttcagacac ttggcgcact tcggtttttc tttggagcac ttgagctttt
     2761 taagtcggca aatatcgcat gcttgttcga tagaagacag tagcttcata gcgctagcgg
     2821 atctgacggt tcactaaacc agctctgctt atatagacct cccaccgtac acgcctaccg
     2881 cccatttgcg tcaatggggc ggagttgtta cgacattttg gaaagtcccg ttgattttgg
     2941 tgccaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca
     3001 aaccgctatc cacgcccatt gatgtactgc caaaaccgca tcaccatggt aatagcgatg
     3061 actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa
     3121 tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac
     3181 acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg
     3241 aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg
     3301 tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata
     3361 tatgggctat gaactaatga ccccgtaatt gattactatt aataactaat gcatggcggt
     3421 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3481 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3541 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3601 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3661 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3721 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3781 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3841 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3901 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3961 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4021 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4081 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4141 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4201 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4261 tgagtaacct gaggctatgg cagggcctgc cgccccgacg ttggctgcga gccctgggcc
     4321 ttcacccgaa cttggggggt ggggtgggga aaaggaagaa acgcgggcgt attggcccca
     4381 atggggtctc ggtggggtat cgacagagtg ccagccctgg gaccgaaccc cgcgtttatg
     4441 aacaaacgac ccaacaccgt gcgttttatt ctgtcttttt attgccgtca tagcgcgggt
     4501 tccttccggt attgtctcct tccgtgtttc agttagcctc cccctagggt gggcgaagaa
     4561 ctccagcatg agatccccgc gctggaggat catccagccg gcgtcccgga aaacgattcc
     4621 gaagcccaac ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg
     4681 cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc agaagaactc gtcaagaagg
     4741 cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg
     4801 tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga
     4861 tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc
     4921 accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc
     4981 atgctcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc
     5041 agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt
     5101 ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca
     5161 tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc
     5221 ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct
     5281 gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttgcagttca
     5341 ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc
     5401 cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc
     5461 ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac
     5521 gatcctcatc ctgtctcttg atcgatcttt gcaaaagcct aggcctccaa aaaagcctcc
     5581 tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa
     5641 aattagtcag ccatggggcg gagaat
//
