LOCUS       sequence-209444-        5805 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses RpBphP1-mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5805)
  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 5805)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-141
FEATURES             Location/Qualifiers
     source          1..5805
                     /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..3145
                     /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"
     primer_bind     complement(3224..3242)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3279..3400
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3290..3309)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3344..3363
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             3799..4593
                     /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(3853..3872)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4463..4482
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4765..5353)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4845..4864)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5469..5752
                     /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 cctgtcacca ttatctgccg cgcttcatcc atttcgagct
     1081 gcgggcgatc tgcgaactgc tcgccgaagc gatcgcgacg cggatcaccg cgcttgagag
     1141 cttcgcgcag agccagtcgg agctgttcgt gcagcggctc gaacagcgca tgatcgaagc
     1201 gatcacccgt gaaggcgatt ggcgcgcagc gattttcgac accagccaat cgatcctgca
     1261 gccgctgcac gccgacggtt gcgcgctggt gtacgaagac cagatcagga ccatcggtga
     1321 cgtaccttcc acgcaggatg ttcgcgagat cgccgggtgg ctcgatcgcc agccacgtgc
     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 agtaatctag agtcgacctg caggcatgca agctgatccg
     3181 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     3241 gcataacccc ttggggcggc cgcttcgagc agacatgata agatacattg atgagtttgg
     3301 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     3361 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     3421 ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta
     3481 caaatgtggt aaaatcgaat tctaatggat cctctttgcg cttgcgtttt cccttgtcca
     3541 gatagcccag tagctgacat tcatccgggg tcagcaccgt ttctgcggac tggctttcta
     3601 cgtgttccgc ttcctttagc agcccttgcg ccctgagtgc ttgcggcagc gtgagcttca
     3661 aaagaattgc cagctggggc gccctctggt aaggttggga agccctgcaa agtaaactgg
     3721 atggctttct tgccgccaag gatctgatgg cgcaggggat caagatctga tcaagagaca
     3781 ggatgacggt cgtttcgcat gcttgaacaa gatggattgc acgcaggttc tccggccgct
     3841 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3901 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3961 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     4021 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4081 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4141 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4201 caccaagcga aacatcgcat cgagcgagca cgcactcgga tggaagccgg tcttgtcgat
     4261 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4321 aaggcgcgta tgccggatgg tgaggatctc gtcgtgactc atggcgatgc ctgcttgccg
     4381 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4441 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4501 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4561 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     4621 accaagcgac gcccaaccgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4681 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4741 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4801 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4861 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4921 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4981 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5041 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5101 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5161 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     5221 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5281 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5341 aaaaggattt caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5401 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5461 tttatagtcc ggaaatacag gaacgcacgc tggatggccc ttcgctggga tggtgaaacc
     5521 atgaaaaatg gcagcttcag tggattaagt gggggtaatg tggcctgtac cctctggttg
     5581 cataggtatt catacggtta aaatttatca ggcgcgattg cggcagtttt tcgggtggtt
     5641 tgttgccatt tttacctgtc tgctgccgtg atcgcgctga acgcgtttta gcggtgcgta
     5701 caattaaggg attatggtaa atccacttac tgtctgccct cgtagccatc gagataaacc
     5761 gcagtactcc ggccacgatg cgtccggcgt agaggatcga gatct
//
