LOCUS       sequence-153237-        4987 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses PpsR2-mVenus-CAAX under CMVd1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4987)
  AUTHORS   Kaberniuk AA, Shemetov AA, Verkhusha VV
  TITLE     A bacterial phytochrome-based optogenetic system controllable with 
            near-infrared light.
  JOURNAL   Nat Methods. 2016 May 9. doi: 10.1038/nmeth.3864.
  PUBMED    27159085
REFERENCE   2  (bases 1 to 4987)
  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 
            Methods. 2016 May 9. doi: 10.1038/nmeth.3864."
COMMENT     Sequence Label: pKA-142
FEATURES             Location/Qualifiers
     source          1..4987
                     /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"
     primer_bind     132..151
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        132..150
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        156..174
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     156..173
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      1661..1670
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1667..1669,1670..1672,1673..2383)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     ERVTAAGITLGMDELYK"
     primer_bind     complement(1712..1733)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1973..1992)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2461..2582
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2472..2491)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2526..2545
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             2981..3775
                     /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="MLEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3035..3054)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3645..3664
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3947..4535)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4027..4046)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4651..4934
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga cgcaaatggg
       61 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
      121 cactagaagc ttaatacgac tcactatagg gtacgattta ggtgacacta tagaataagg
      181 agcgatcgcc atggctagcg cgtcaaagtc cgttcatgcc gacatcaccc ttctgctcga
      241 tatggagggt gtgattcgcg aagccaccct gtctccgacg atggcggccg agagcgtgga
      301 cggttggctg gggcgtcgct ggagcgacat cgccggcgcc gaaggcggcg acaaggttcg
      361 ccgcatggtc gaagacgccc gccgcagcgg catctcggct ttccgccaga tcaatcagcc
      421 tttcccgagc ggcgtcgaaa tcccgatcga attcaccacg atgctgctgg gcgaccgcac
      481 cggcatgatc gcggtcggca agaacatgca ggcggtcacc gagctgcatt cccggctgat
      541 cgctgcgcag caggcgatgg agcgcgacta ttggcggttg cgtgaattgg agactcgcta
      601 ccgcctggtg ttcgacgctg ccgccgatgc ggtgatgatc gtctccgccg gcgacatgcg
      661 catcgtcgaa gccaaccggg cggcggtgaa tgcgatcagc cgcgtcgagc gcggcaatga
      721 cgaccttgcg gggcgtgatt tcctcgccga agtggcggct gccgatcgcg atgcggtgcg
      781 cgacatgctg gcccaggtgc gtcagcgcgg caccgcactc agcgtcctcg ttcatctcgg
      841 ccgttacgac cgcgcctgga tgctgcgcgg ttcgctgatg tcgtccgagc gtcgtcaggt
      901 tttcctgctg cacttcaccc cggtgaccac gactcccgcg atcgacgacg tcgacgatga
      961 tgccgtgctg cgcgggctga tcgatcgcat tcccgacggg ttcgtcgcac tggattcgga
     1021 aggcgtcgtt cgtcacgcca accaggcgtt tctcgatctg gtccagatcg gctccaagcc
     1081 tgcggcggtc ggacgatcgc tgggcgtctg gatgggtcgt ccgggcgccg atctgtccag
     1141 cttgctgacg ctgctgcggc gctacaagac ggtgcggctg ttccaaacga cgatccgcgg
     1201 cgagctcggc accgagactg aagtcgaggt ctcggccgtc gacggcgagg acgaccaata
     1261 catcggcgtt ctgatgcgca atgtcgcgcg acgcctcgac gctgcggacg accacgatgc
     1321 cttgcgtcag gcgctcggcc cgatcagcaa gcagctcggg cgatcctcgc tgcgcaagct
     1381 ggtgaagaac gccgtgagca ttgtcgagca gcactacgtg aaggaagcgc tgttgcgatc
     1441 caagggcaat cgcacggcaa ctgccgaact gctcggattg agccggcaga gcctttatgc
     1501 aaaactcaac tcctacggct tcgacgacaa aggtgtcgtt gcttctgctg ccgacggtgc
     1561 agagggcgcc tcagacgacg cagaggataa gcttagtgct ggtggtagtg ctggtggtag
     1621 tgctggtggt agtgctggtg gtagtgctgg tggaccggtc gccaccatgg tgagcaaggg
     1681 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     1741 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     1801 gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     1861 gggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
     1921 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     1981 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     2041 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     2101 ctacaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa
     2161 cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca
     2221 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc taagctacca
     2281 gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggaacgcgt
     2341 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aaggctagcg gtaaaaagaa
     2401 gaaaaagaag tcaaagacaa agtgtgtaat tatgtaagcg gccgcttcga gcagacatga
     2461 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     2521 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     2581 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
     2641 ttttaagcaa gtaaaacctc tacaaatgtg gtaaaatcga attctaatgg atcctctttg
     2701 cgcttgcgtt ttcccttgtc cagatagccc agtagctgac attcatccgg ggtcagcacc
     2761 gtttctgcgg actggctttc tacgtgttcc gcttccttta gcagcccttg cgccctgagt
     2821 gcttgcggca gcgtgagctt caaaagaatt gccagctggg gcgccctctg gtaaggttgg
     2881 gaagccctgc aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg
     2941 atcaagatct gatcaagaga caggatgacg gtcgtttcgc atgcttgaac aagatggatt
     3001 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3061 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3121 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     3181 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3241 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3301 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3361 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgcactcg
     3421 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     3481 agccgaactg ttcgccaggc tcaaggcgcg tatgccggat ggtgaggatc tcgtcgtgac
     3541 tcatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     3601 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     3661 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     3721 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     3781 actctggggt tcgaaatgac cgaccaagcg acgcccaacc ggtatcagct cactcaaagg
     3841 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3901 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3961 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4021 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4081 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4141 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4201 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4261 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4321 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4381 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4441 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4501 agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg
     4561 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4621 aaaggatctt cacctagatc cttttatagt ccggaaatac aggaacgcac gctggatggc
     4681 ccttcgctgg gatggtgaaa ccatgaaaaa tggcagcttc agtggattaa gtgggggtaa
     4741 tgtggcctgt accctctggt tgcataggta ttcatacggt taaaatttat caggcgcgat
     4801 tgcggcagtt tttcgggtgg tttgttgcca tttttacctg tctgctgccg tgatcgcgct
     4861 gaacgcgttt tagcggtgcg tacaattaag ggattatggt aaatccactt actgtctgcc
     4921 ctcgtagcca tcgagataaa ccgcagtact ccggccacga tgcgtccggc gtagaggatc
     4981 gagatct
//
