pUBC-mCherry-1POK

LOCUS       pUBC-mCherry-1POK                6342 bp DNA     circular SYN 12-MAY-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6342)
  AUTHORS   Ruizhao Wang
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6342
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     56..101
                     /label=pUBC-mCherry-F
     regulatory      77..86
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             83..790
                     /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(767..814)
                     /label=mCherry-Flag-R
     primer_bind     791..834
                     /label=Flag-1POK-F
     CDS             791..814
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             815..850
                     /codon_start=1
                     /label=Linker12
                     /translation="GGSGGTGGSGGT"
     CDS             851..2017
                     /codon_start=1
                     /label=1P0K(E239Y)
                     /translation="IDYTAAGFTLLQGAHLYAPEDRGICDVLVANGKIIAVASNIPSDI
                     VPNCTVVDLSGQILCPGFIDQHVHLIGGGGEAGPTTRTPEVALSRLTEAGVTSVVGLLG
                     TDSISRHPESLLAKTRALNEEGISAWMLTGAYHVPSRTITGSVEKDVAIIDRVIGVKCA
                     ISDHRSAAPDVYHLANMAAESRVGGLLGGKPGVTVFHMGDSKKALQPIYDLLENCDVPI
                     SKLLPTHVNRNVPLFYQALEFARKGGTIDITSSIDEPVAPAEGIARAVQAGIPLARVTL
                     SSDGNGSQPFFDDEGNLTHIGVAGFETLLETVQVLVKDYDFSISDALRPLTSSVAGFLN
                     LTGKGEILPGNDADLLVMTPELRIEQVYARGKLMVKDGKACVKGTFETA"
     primer_bind     complement(2114..2137)
                     /label=pCAG-R
     polyA_signal    2171..2226
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2587..2603)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2611..2627
                     /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(2635..2665)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     2640..2661
                     /label=lac-pro-F
     protein_bind    2680..2701
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2760..2955
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        2785..2955
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     primer_bind     2801..2813
                     /label=mCherry-Flag-R
     rep_origin      2806..2941
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2961..3095
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3334..3922)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3338..3357)
                     /label=pUBC-primer-1
     CDS             complement(4093..4953)
                     /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     complement(4124..4145)
                     /label=Amp-F
     primer_bind     4867..4887
                     /label=Amp-R-seq
     promoter        complement(4954..5057)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5130..5529
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(5257..5279)
                     /label=hUBC-pro-R
     primer_bind     5725..5748
                     /label=pUBC-seq-2
     promoter        5816..6342
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6179..6198
                     /label=UBC-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
ORIGIN
        1 gaagcttggg ctgcaggtcg agctcaagct tcgaattctg cagtcgacgg taccgcgggc
       61 ccgggatcca ccggtcgcca ccatggtgag caagggcgag gaggataaca tggccatcat
      121 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga
      181 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
      241 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg
      301 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc
      361 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
      421 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
      481 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga
      541 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa
      601 ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca
      661 gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta
      721 caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga
      781 gctgtacaag gattacaagg acgacgatga caagggcggc agcggcggca ccggcggcag
      841 cggcggcacc atcgactaca ccgccgccgg cttcaccctg ctgcagggcg cccacctgta
      901 cgcccccgag gacaggggca tctgcgacgt gctggtggcc aacggcaaga tcatcgccgt
      961 ggccagcaac atccccagcg acatcgtgcc caactgcacc gtggtggacc tgagcggcca
     1021 gatcctgtgc cccggcttca tcgaccagca cgtgcacctg atcggcggcg gcggcgaggc
     1081 cggccccacc accaggaccc ccgaggtggc cctgagcagg ctgaccgagg ccggcgtgac
     1141 cagcgtggtg ggcctgctgg gcaccgacag catcagcagg caccccgaga gcctgctggc
     1201 caagaccagg gccctgaacg aggagggcat cagcgcctgg atgctgaccg gcgcctacca
     1261 cgtgcccagc aggaccatca ccggcagcgt ggagaaggac gtggccatca tcgacagggt
     1321 gatcggcgtg aagtgcgcca tcagcgacca caggagcgcc gcccccgacg tgtaccacct
     1381 ggccaacatg gccgccgaga gcagggtggg cggcctgctg ggcggcaagc ccggcgtgac
     1441 cgtgttccac atgggcgaca gcaagaaggc cctgcagccc atctacgacc tgctggagaa
     1501 ctgcgacgtg cccatcagca agctgctgcc cacccacgtg aacaggaacg tgcccctgtt
     1561 ctaccaggcc ctggagttcg ccaggaaggg cggcaccatc gacatcacca gcagcatcga
     1621 cgagcccgtg gcccccgccg agggcatcgc cagggccgtg caggccggca tccccctggc
     1681 cagggtgacc ctgagcagcg acggcaacgg cagccagccc ttcttcgacg acgagggcaa
     1741 cctgacccac atcggcgtgg ccggcttcga gaccctgctg gagaccgtgc aggtgctggt
     1801 gaaggactac gacttcagca tcagcgacgc cctgaggccc ctgaccagca gcgtggccgg
     1861 cttcctgaac ctgaccggca agggcgagat cctgcccggc aacgacgccg acctgctggt
     1921 gatgaccccc gagctgagga tcgagcaggt gtacgccagg ggcaagctga tggtgaagga
     1981 cggcaaggcc tgcgtgaagg gcaccttcga gacggcctga gcggccgcac tcctcaggtg
     2041 caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac
     2101 tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg
     2161 acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt
     2221 ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg
     2281 gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa
     2341 agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc
     2401 ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc
     2461 ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca
     2521 gtcatagctg tccctcttct cttatggaga tccctcgacc tgcagcccaa gcttggcgta
     2581 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     2641 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     2701 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg
     2761 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     2821 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     2881 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     2941 ctaggctttt gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca
     3001 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     3061 ccaaactcat caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg
     3121 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3181 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3241 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3301 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3361 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3421 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3481 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3541 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3601 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3661 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3721 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3781 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3841 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3901 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3961 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4021 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4081 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4141 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4201 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4261 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4321 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4381 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4441 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4501 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4561 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4621 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4681 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4741 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4801 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4861 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4921 aataagggcg acacggaaat gttgaatact catactcttc ctttttcata ttattgaagc
     4981 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5041 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgggtcgac attgattatt
     5101 gactagtaac ccgtgtcggc tccagatctg gcctccgcgc cgggttttgg cgcctcccgc
     5161 gggcgccccc ctcctcacgg cgagcgctgc cacgtcagac gaagggcgca gcgagcgtcc
     5221 tgatccttcc gcccggacgc tcaggacagc ggcccgctgc tcataagact cggccttaga
     5281 accccagtat cagcagaagg acattttagg acgggacttg ggtgactcta gggcactggt
     5341 tttctttcca gagagcggaa caggcgagga aaagtagtcc cttctcggcg attctgcgga
     5401 gggatctccg tggggcggtg aacgccgatg attatataag gacgcgccgg gtgtggcaca
     5461 gctagttccg tcgcagccgg gatttgggtc gcggttcttg tttgtggatc gctgtgatcg
     5521 tcacttggtg agtagcgggc tgctgggctg gccggggctt tcgtggccgc cgggccgctc
     5581 ggtgggacgg aagcgtgtgg agagaccgcc aagggctgta gtctgggtcc gcgagcaagg
     5641 ttgccctgaa ctgggggttg ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg
     5701 aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg
     5761 ggcggcaaga acccaaggtc ttgaggcctt cgctaatgcg ggaaagctct tattcgggtg
     5821 agatgggctg gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact
     5881 cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt
     5941 acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat
     6001 gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag
     6061 ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag
     6121 ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg
     6181 aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag
     6241 gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gactagtaaa
     6301 ttgtccgcta aattctggcc gtttttggct tttttgttag ac
//