LOCUS       sequence-460768-        6400 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 6400)
  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 6400)
  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: cpLOV2-Gal4
FEATURES             Location/Qualifiers
     source          1..6400
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(20..39)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        211..590
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        591..794
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     745..765
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     839..858
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        839..857
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1354..1374
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1393..1830
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="KLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTKR
                     SPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNKD
                     AVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     primer_bind     complement(1967..1984)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1973..2197
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2243..2671
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2330..2349)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2540..2561
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2680..2700)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2685..3014
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2865..3000
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2927..2946
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3081..3875
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3135..3154)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3745..3764
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4049..4170
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4086..4105)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4140..4159
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4219..4235)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4219..4235)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4232..4254)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4243..4259
                     /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(4267..4273,4274..4291,4292..4297))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4312..4333
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4450..4467)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4621..5209)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4701..4720)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5380..6171,6172..6240))
                     /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     6003..6022
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6241..6345)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
       61 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
      121 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
      181 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
      241 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      301 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      361 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      421 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      481 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      541 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      601 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      661 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      721 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      781 atataagcag agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta
      841 atacgactca ctatagggag acccaagctg gctagcatga ctgaacacgt cagagatgcg
      901 gcggagcgag aaggagtgat gctgattaag aaaacagcag agaatatcga cgaagcggcg
      961 aaagagttgg gtgggggatc cggtggctcc ggaggtggtc ttgctacaac tttggagcgg
     1021 attgagaaga atttcgtcat aactgatccc aggttgcctg ataatcctat aatattcgcc
     1081 tccgatagtt ttcttcaact tacggagtac tccagagagg agatactggg aaggaattgc
     1141 agattcctcc aaggtccaga aactgataga gctactgtga ggaagatacg ggatgccatt
     1201 gacaatcaaa ctgaagtgac agtgcaactt atcaattaca ccaagtcagg gaaaaaattc
     1261 tggaacctgt tccatttgca acccatgagg gaccaaaagg gcgatgtgca atatttcata
     1321 ggcgttcaac ttgatggtgg aggtggaggt agccccaaaa agaagcgaaa agtaggcggc
     1381 agtggcggga gtaagctcct cagttctata gaacaggcgt gcgatatctg tcgactgaaa
     1441 aaactgaagt gtagtaaaga aaagccaaag tgtgcaaagt gtttgaagaa caattgggaa
     1501 tgcagatatt caccgaagac caaacggagc cctcttactc gcgcccattt gacggaagtt
     1561 gaatccaggc tcgaaaggtt ggagcaactc tttcttttga ttttcccgag ggaagatctt
     1621 gacatgattt tgaaaatgga ttcattgcaa gatattaaag cactgcttac ggggttgttt
     1681 gtccaagaca acgtgaacaa ggacgctgtc acagatcgcc tggctagtgt tgaaacagat
     1741 atgccgttga ctttgaggca acacaggatt agcgcaacct cctcttctga agaatcctct
     1801 aataaagggc agcggcaatt gacggtcagc ggcggctccg gtggagggtc cgggggaagt
     1861 tcatgactcg agcggccgcc actgtgctgg atatctgcag aattccacca cactggacta
     1921 gtggatccga gctcggtacc aagcttaagt ttaaaccgct gatcagcctc gactgtgcct
     1981 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     2041 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2101 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     2161 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     2221 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2281 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2341 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2401 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     2461 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2521 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     2581 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     2641 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     2701 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2761 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2821 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     2881 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     2941 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3001 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     3061 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3121 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3181 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3241 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     3301 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     3361 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     3421 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     3481 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     3541 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     3601 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     3661 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     3721 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     3781 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     3841 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     3901 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     3961 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4021 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     4081 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4141 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     4201 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4261 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4321 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4381 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4441 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4501 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4561 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4621 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4681 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4741 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4801 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4861 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4921 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4981 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5041 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5101 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5161 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5221 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5281 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5341 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5401 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5461 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5521 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5581 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5641 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5701 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5761 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5821 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5881 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5941 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6001 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6061 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6121 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6181 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6241 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6301 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6361 agtgccacct gacgtcgacg gatcgggaga tctcccgatc
//
