LOCUS       sequence-210752-        6498 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent PKCZ.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6498)
  TITLE     Expression of wt and mt tagged-human genes and oncogenes
REFERENCE   2  (bases 1 to 6498)
  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"
COMMENT     Sequence Label: mCherry-PKCZ
FEATURES             Location/Qualifiers
     source          1..6498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2387..2396
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2393..3103
                     /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(2538..2556)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2806..2825
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3226..3347
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3263..3282)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3317..3336
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3354..3809)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3491..3512)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3703..3722
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3836..3940
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3942..4299
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3942..4133
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3965..3985)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4150..4285
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4334..5128
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4334..5128
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4388..4407)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4998..5017
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5316..5335)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5360..5407
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5736..6324
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6225..6244
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attcatgccc agcaggaccg gccccaagat
      601 ggaagggagc ggcggccgcg tccgcctcaa ggcgcattac gggggggaca tcttcatcac
      661 cagcgtggac gccgccacga ccttcgagga gctctgtgag gaagtgagag acatgtgtcg
      721 tctgcaccag cagcacccgc tcaccctcaa gtgggtggac agcgaaggtg acccttgcac
      781 ggtgtcctcc cagatggagc tggaagaggc tttccgcctg gcccgtcagt gcagggatga
      841 aggcctcatc attcatgttt tcccgagcac ccctgagcag cctggcctgc catgtccggg
      901 agaagacaaa tctatctacc gccggggagc cagaagatgg aggaagctgt accgtgccaa
      961 cggccacctc ttccaagcca agcgctttaa caggagagcg tactgcggtc agtgcagcga
     1021 gaggatatgg ggcctcgcga ggcaaggcta caggtgcatc aactgcaaac tgctggtcca
     1081 taagcgctgc cacggcctcg tcccgctgac ctgcaggaag catatggatt ctgtcatgcc
     1141 ttcccaagag cctccagtag acgacaagaa cgaggacgcc gaccttcctt ccgaggagac
     1201 agatggaatt gcttacattt cctcatcccg gaagcatgac agcattaaag acgactcgga
     1261 ggaccttaag ccagttatcg atgggatgga tggaatcaaa atctctcagg ggcttgggct
     1321 gcaggacttt gacctaatca gagtcatcgg gcgcgggagc tacgccaagg ttctcctggt
     1381 gcggttgaag aagaatgacc aaatttacgc catgaaagtg gtgaagaaag agctggtgca
     1441 tgatgacgag gatattgact gggtacagac agagaagcac gtgtttgagc aggcatccag
     1501 caaccccttc ctggtcggat tacactcctg cttccagacg acaagtcggt tgttcctggt
     1561 cattgagtac gtcaacggcg gggacctgat gttccacatg cagaggcaga ggaagctccc
     1621 tgaggagcac gccaggttct acgcggccga gatctgcatc gccctcaact tcctgcacga
     1681 gagggggatc atctacaggg acctgaagct ggacaacgtc ctcctggatg cggacgggca
     1741 catcaagctc acagactacg gcatgtgcaa ggaaggcctg ggccctggtg acacaacgag
     1801 cactttctgc ggaaccccga attacatcgc ccccgaaatc ctgcggggag aggagtacgg
     1861 gttcagcgtg gactggtggg cgctgggagt cctcatgttt gagatgatgg ccgggcgctc
     1921 cccgttcgac atcatcaccg acaacccgga catgaacaca gaggactacc ttttccaagt
     1981 gatcctggag aagcccatcc ggatcccccg gttcctgtcc gtcaaagcct cccatgtttt
     2041 aaaaggattt ttaaataagg accccaaaga gaggctcggc tgccggccac agactggatt
     2101 ttctgacatc aagtcccacg cgttcttccg cagcatagac tgggacttgc tggagaagaa
     2161 gcaggcgctc cctccattcc agccacagat cacagacgac tacggtctgg acaactttga
     2221 cacacagttc accagcgagc ccgtgcagct gaccccagac gatgaggatg ccataaagag
     2281 gatcgaccag tcagagttcg aaggctttga gtatatcaac ccattattgc tgtccaccga
     2341 ggagtcggtg cagtcgacgg taccgcgggc ccgggatcca ccggtcgcca ccatggtgag
     2401 caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat
     2461 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
     2521 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg
     2581 ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga
     2641 catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     2701 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt
     2761 catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa
     2821 gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa
     2881 gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa
     2941 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa
     3001 gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga
     3061 gggccgccac tccaccggcg gcatggacga gctgtacaag taaagcggcc gcgactctag
     3121 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     3181 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     3241 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3301 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     3361 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     3421 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     3481 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     3541 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     3601 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     3661 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     3721 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     3781 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     3841 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3901 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     3961 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4021 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4081 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4141 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4201 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     4261 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     4321 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4381 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4441 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4501 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4561 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4621 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4681 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4741 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4801 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4861 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4921 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4981 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5041 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5101 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     5161 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5221 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5281 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     5341 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     5401 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     5461 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     5521 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     5581 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5641 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5701 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5761 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5821 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5881 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5941 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6001 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6061 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6121 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6181 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6241 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6301 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6361 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     6421 cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag
     6481 cccatatatg gagttccg
//
