LOCUS       sequence-459787-        5354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses components of the LOOMINA optogenetic transcriptional 
            control system..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5354)
  AUTHORS   Muench P, Fiumara M, Southern N, Coda D, Aschenbrenner S, Correia B,
            Graff J, Niopek D, Mathony J
  TITLE     A modular toolbox for the optogenetic deactivation of transcription.
  JOURNAL   Nucleic Acids Res. 2024 Dec 16:gkae1237. doi: 10.1093/nar/gkae1237.
  PUBMED    39676667
REFERENCE   2  (bases 1 to 5354)
  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: "Nucleic 
            Acids Res. 2024 Dec 16:gkae1237. doi: 10.1093/nar/gkae1237."
COMMENT     Sequence Label: CMV_Zdk3-VPR
FEATURES             Location/Qualifiers
     source          1..5354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             7..24
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(47..64)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    53..277
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(348..364)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(348..364)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(361..383)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    372..388
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(396..402,403..420,421..426))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    441..462
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(579..596)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(750..1338)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(830..849)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1509..2300,2301..2369))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2132..2151
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2370..2474)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2553..2572)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2744..3123
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3124..3327
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3278..3298
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3369..3388
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3369..3387
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3416..3425
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3434..3454
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3716..3865
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             4334..4690
                     /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
                     EDFSSIADMDFSALL"
     CDS             4709..5278
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     CDS             5300..5320
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 accggtcatc atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct
       61 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
      121 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
      181 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
      241 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
      301 tggggctcga taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
      361 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
      421 tgtaaagcct agggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
      481 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
      541 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
      601 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
      661 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
      721 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
      781 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
      841 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
      901 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
      961 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     1021 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     1081 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     1141 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     1201 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     1261 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     1321 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cactcagtgg
     1381 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     1441 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     1501 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     1561 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     1621 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     1681 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     1741 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     1801 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     1861 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     1921 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     1981 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     2041 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     2101 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     2161 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     2221 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     2281 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     2341 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     2401 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     2461 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     2521 cgatctcccg atcccctagg gtcgactctc agtacaatct gctctgatgc cgcatagtta
     2581 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     2641 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
     2701 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact
     2761 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat tgagttccgc
     2821 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     2881 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     2941 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     3001 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     3061 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     3121 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     3181 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     3241 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     3301 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagagatcc
     3361 gcggccgcta atacgactca ctatagggag agccgccacc gtgtcgtgac gtacggccac
     3421 catgggtagc ggacctaaga aaaagaggaa ggtggcggcc gctggaggtt ctggaagtgg
     3481 atccatggtg gataacaaat tcaataaaga agtgctggtg gcccggcagg aaatctattg
     3541 gctgccaaac ctgaattggg agcagaagtt tgccttcatc agtagcctga ctaatgatcc
     3601 atctcagagc gccaatctgc tggccgaagc caaaaaactg aacggtgccc agcccccaaa
     3661 aggaggaggt ggaagcggag gcggcggaag tgcggccgct ggatccggac gggctgacgc
     3721 attggacgat tttgatctgg atatgctggg aagtgacgcc ctcgatgatt ttgaccttga
     3781 catgcttggt tcggatgccc ttgatgactt tgacctcgac atgctcggca gtgacgccct
     3841 tgatgatttc gacctggaca tgctgattaa cactagtggc ggctctggcg gcggctccgg
     3901 aggctctagc cagtacctgc ccgacaccga cgacaggcac cggatcgagg aaaagcggaa
     3961 gcggacctac gagacattca agagcatcat gaagaagtcc cccttcagcg gccccaccga
     4021 ccctagacct ccacctagaa gaatcgccgt gcccagcaga tccagcgcca gcgtgccaaa
     4081 acctgccccc cagccttacc ccttcaccag cagcctgagc accatcaact acgacgagtt
     4141 ccctaccatg gtgttcccca gcggccagat ctctcaggcc tctgctctgg ctccagcccc
     4201 tcctcaggtg ctgcctcagg ctcctgctcc tgcaccagct ccagccatgg tgtctgcact
     4261 ggctcaggca ccagcacccg tgcctgtgct ggctcctgga cctccacagg ctgtggctcc
     4321 accagcccct aaacctacac aggccggcga gggcacactg tctgaagctc tgctgcagct
     4381 gcagttcgac gacgaggatc tgggagccct gctgggaaac agcaccgatc ctgccgtgtt
     4441 caccgacctg gccagcgtgg acaacagcga gttccagcag ctgctgaacc agggcatccc
     4501 tgtggcccct cacaccaccg agcccatgct gatggaatac cccgaggcca tcacccggct
     4561 cgtgacaggc gctcagaggc ctcctgatcc agctcctgcc cctctgggag caccaggcct
     4621 gcctaatgga ctgctgtctg gcgacgagga cttcagctct atcgccgata tggatttctc
     4681 agccttgctg ggctctggca gcggcagccg ggattccagg gaagggatgt ttttgccgaa
     4741 gcctgaggcc ggctccgcta ttagtgacgt gtttgagggc cgcgaggtgt gccagccaaa
     4801 acgaatccgg ccatttcatc ctccaggaag tccatgggcc aaccgcccac tccccgccag
     4861 cctcgcacca acaccaaccg gtccagtaca tgagccagtc gggtcactga ccccggcacc
     4921 agtccctcag ccactggacc cagcgcccgc agtgactccc gaggccagtc acctgttgga
     4981 ggaccccgat gaagagacga gccaggctgt caaagccctt cgggagatgg ccgatactgt
     5041 gattccccag aaggaagagg ctgcaatctg tggccaaatg gacctttccc atccgccccc
     5101 aaggggccat ctggatgagc tgacaaccac acttgagtcc atgaccgagg atctgaacct
     5161 ggactcaccc ctgaccccgg aattgaacga gattctggat accttcctga acgacgagtg
     5221 cctcttgcat gccatgcata tcagcacagg actgtccatc ttcgacacat ctctgtttgg
     5281 ttctggaagt ggatccagcc ccaagaagaa gagaaaggtg gaggccagct aaagcgaagt
     5341 caatcccaga attc
//
