LOCUS       sequence-155710-        6808 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses RpBphP1-mCherry-Intersectin1 DHPH domain under CMVd1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6808)
  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 6808)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-144
FEATURES             Location/Qualifiers
     source          1..6808
                     /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"
     CDS             2435..3142
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2580..2598)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2848..2867
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4282..4403
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4293..4312)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4347..4366
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             4802..5596
                     /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(4856..4875)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5466..5485
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(5768..6356)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5848..5867)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6472..6755
                     /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 tggcaggtca tgcctctggc agccccgcat tcgggaccgc
      241 cgatctttcg aattgcgaac gtgaagagat ccacctcgcc ggctcgatcc agccgcatgg
      301 cgcgcttctg gtcgtcagcg agccggatca tcgcatcatc caggccagcg ccaacgccgc
      361 ggaatttctg aatctcggaa gcgtgctcgg cgttccgctc gccgagatcg acggcgatct
      421 gttgatcaag atcctgccgc atctcgatcc caccgccgaa ggcatgccgg tcgcggtgcg
      481 ctgccggatc ggcaatccct ccacggagta cgacggtctg atgcatcggc ctccggaagg
      541 cgggctgatc atcgagctcg aacgtgccgg cccgccgatc gatctgtccg gcacgctggc
      601 gccggcgctg gagcggatcc gcacggcggg ctcgctgcgc gcgctgtgcg atgacaccgc
      661 gctgctgttt cagcagtgca ccggctacga ccgggtgatg gtgtatcgct tcgacgagca
      721 gggccacggc gaagtgttct ccgagcgcca cgtgcccggg ctcgaatcct atttcggcaa
      781 ccgctatccg tcgtcggaca ttccgcagat ggcgcggcgg ctgtacgagc ggcagcgcgt
      841 ccgcgtgctg gtcgacgtca gctatcagcc ggtgccgctg gagccgcggc tgtcgccgct
      901 gaccgggcgc gatctcgaca tgtcgggctg cttcctgcgc tcgatgtcgc cgatccatct
      961 gcagtacctg aagaacatgg gcgtgcgcgc caccctggtg gtgtcgctgg tggtcggcgg
     1021 caagctgtgg ggcctggttg cctgtcatca ttatctgccg cgcttcatgc atttcgagct
     1081 gcgggcgatc tgcgaactgc tcgccgaagc gatcgcgacg cggatcaccg cgcttgagag
     1141 cttcgcgcag agccagtcgg agctgttcgt gcagcggctc gaacagcgca tgatcgaagc
     1201 gattacccgt gaaggcgatt ggcgcgcagc gattttcgac accagccaat cgatcctgca
     1261 gccgctgcac gccgccggtt gcgcgctggt gtacgaagac cagatcagga ccatcggcga
     1321 cgtgccttcc acgcaggatg tgcgcgagat cgccgggtgg ctcgatcgcc agccgcgcgc
     1381 ggcggtgacc tcgaccgcgt cgctcggtct cgacgtgccg gagctcgcgc atctgacgcg
     1441 gatggcgagc ggcgtggtcg cggcgccgat ttcggatcat cgcggcgagt ttctgatgtg
     1501 gttccgcccc gagcgcgtcc acaccgttac ctggggcggc gatccgaaga agccgttcac
     1561 gatgggcgat acaccggcgg atctgtcgcc gcggcgctcc ttcgccaaat ggcatcaggt
     1621 tgtcgaaggc acgtccgatc cgtggacggc cgccgatctc gccgcggctc gcaccatcgg
     1681 tcagaccgtc gccgacatcg tgctgcaatt ccgcgcggtg cggacactga tcgcccgcga
     1741 acagtacgaa cagttttcgt cccaggtgca cgcttcgatg cagccggtgc tgatcaccga
     1801 cgccgaaggc cgcatcctgc tgatgaacga ctcgttccgc gacatgttgc cggcggggtc
     1861 gccatccgcc gtccatctcg acgatctcgc cgggttcttc gtcgaatcga acgatttcct
     1921 gcgcaacgtc gccgaactga tcgatcacgg ccgcgggtgg cgcggcgaag ttctgctgcg
     1981 cggcgcaggt aatcgcccgt tgccgctggc agtgcgcgcc gatccggtga cgcgcacgga
     2041 ggaccagtcg ctcggcttcg tgctgatctt cagcgacgct accgatcgtc gcaccgcaga
     2101 tgccgcacgc acgcgtttcc aggaaggcat tcttgccagc gcacgtcccg gcgtgcggct
     2161 cgactccaag tccgacctct tgcacgagaa gctgctgtcc gcgctggtcg agaacgcgca
     2221 gcttgccgca ttggaaatta cttacggcgt cgagaccgga cgcatcgccg agctgctcga
     2281 aggcgttcgc cagtcgatgc tgcgcaccgc cgaagtgctc ggccatctgg tgcagcacgc
     2341 ggcgcgcacg gccggcagcg acagctcgag caatggctcg cagaacaaga aggaattcga
     2401 tagtgctggt agtgctggta gtgctggtac tagtatggtg agcaagggcg aggaggataa
     2461 catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg
     2521 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     2581 caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca
     2641 gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa
     2701 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
     2761 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
     2821 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga
     2881 ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag
     2941 gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa
     3001 gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca
     3061 caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg
     3121 cggcatggac gagctgtaca agagtgctgg tgggtccgcc ggcgggagtg ctggcggtag
     3181 tgccggcggt agttctagag atatgttgac cccaactgaa agaaagcgac aaggatacat
     3241 ccacgagctc attgtcaccg aggagaacta tgtgaatgac ctgcagctgg tcacagagat
     3301 ttttcaaaaa cccctgatgg agtctgagct gctgacagaa aaagaggttg ctatgatttt
     3361 tgtgaactgg aaggagctga ttatgtgtaa tatcaaacta ctaaaagcgc tgagagtccg
     3421 caagaagatg tccggggaga agatgcctgt gaagatgatt ggagacatcc tgagcgcaca
     3481 gctgccgcac atgcagccct acatccgctt ctgcagccgc cagctcaacg gggctgccct
     3541 gatccagcag aagacggacg aggccccaga cttcaaggag ttcgtcaaaa gattggcaat
     3601 ggatcctcgg tgtaaaggga tgccactctc tagttttata ctgaagccta tgcaacgggt
     3661 aacaagatac ccactgatca ttaaaaatat cctggaaaac acccctgaaa accacccgga
     3721 ccacagccac ttgaagcacg ccctggagaa ggcggaagag ctctgttccc aggtgaacga
     3781 aggggtgcgg gagaaggaga actctgaccg gctggagtgg atccaggccc acgtgcagtg
     3841 tgaaggcctg tctgagcaac ttgtgttcaa ttcagtgacc aattgcttgg ggccgcgcaa
     3901 atttctgcac agtgggaagc tctacaaggc caagagcaac aaggagctgt atggcttcct
     3961 tttcaacgac ttcctcctgc tgactcagat cacgaagcct ttggggtctt ctggcaccga
     4021 caaagtcttc agccccaaat caaacctgca gtataaaatg tataaaacac ctattttcct
     4081 aaatgaggtt ctagtaaaat tacccaccga cccttctgga gacgagccca tcttccacat
     4141 ctcccacatt gaccgcgtct atactctccg agcagaaagc ataaatgaaa ggactgcctg
     4201 ggtgcagaaa atcaaagctg cttctgaact ctacatagag actgagaaaa agaagtaagc
     4261 ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     4321 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     4381 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     4441 agggggagat gtgggaggtt tttttaagca agtaaaacct ctacaaatgt ggtaaaatcg
     4501 aattctaatg gatcctcttt gcgcttgcgt tttcccttgt ccagatagcc cagtagctga
     4561 cattcatccg gggtcagcac cgtttctgcg gactggcttt ctacgtgttc cgcttccttt
     4621 agcagccctt gcgccctgag tgcttgcggc agcgtgagct tcaaaagaat tgccagctgg
     4681 ggcgccctct ggtaaggttg ggaagccctg caaagtaaac tggatggctt tcttgccgcc
     4741 aaggatctga tggcgcaggg gatcaagatc tgatcaagag acaggatgac ggtcgtttcg
     4801 catgcttgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4861 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4921 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4981 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     5041 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     5101 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     5161 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     5221 catcgagcga gcacgcactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     5281 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gtatgccgga
     5341 tggtgaggat ctcgtcgtga ctcatggcga tgcctgcttg ccgaatatca tggtggaaaa
     5401 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     5461 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5521 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     5581 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     5641 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5701 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5761 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5821 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5881 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5941 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6001 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6061 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6121 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6181 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     6241 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6301 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tttcaagaag
     6361 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6421 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttatag tccggaaata
     6481 caggaacgca cgctggatgg cccttcgctg ggatggtgaa accatgaaaa atggcagctt
     6541 cagtggatta agtgggggta atgtggcctg taccctctgg ttgcataggt attcatacgg
     6601 ttaaaattta tcaggcgcga ttgcggcagt ttttcgggtg gtttgttgcc atttttacct
     6661 gtctgctgcc gtgatcgcgc tgaacgcgtt ttagcggtgc gtacaattaa gggattatgg
     6721 taaatccact tactgtctgc cctcgtagcc atcgagataa accgcagtac tccggccacg
     6781 atgcgtccgg cgtagaggat cgagatct
//
