pUBC-mVenus-1POK

LOCUS       pUBC-mVenus-1POK               6354 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 6354)
  AUTHORS   Ruizhao Wang
  TITLE     Direct Submission
  JOURNAL   Exported Jul 13, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      77..86
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             83..799
                     /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
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(128..149)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(389..408)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     736..757
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     778..826
                     /label=mVenus-1POK-F
     CDS             800..826
                     /codon_start=1
                     /label=HA tag
                     /translation="YPYDVPDYA"
     CDS             827..862
                     /codon_start=1
                     /label=Linker12
                     /translation="GGSGGTGGSGGT"
     CDS             863..2029
                     /codon_start=1
                     /label=1P0K(E239Y)
                     /translation="IDYTAAGFTLLQGAHLYAPEDRGICDVLVANGKIIAVASNIPSDI
                     VPNCTVVDLSGQILCPGFIDQHVHLIGGGGEAGPTTRTPEVALSRLTEAGVTSVVGLLG
                     TDSISRHPESLLAKTRALNEEGISAWMLTGAYHVPSRTITGSVEKDVAIIDRVIGVKCA
                     ISDHRSAAPDVYHLANMAAESRVGGLLGGKPGVTVFHMGDSKKALQPIYDLLENCDVPI
                     SKLLPTHVNRNVPLFYQALEFARKGGTIDITSSIDEPVAPAEGIARAVQAGIPLARVTL
                     SSDGNGSQPFFDDEGNLTHIGVAGFETLLETVQVLVKDYDFSISDALRPLTSSVAGFLN
                     LTGKGEILPGNDADLLVMTPELRIEQVYARGKLMVKDGKACVKGTFETA"
     polyA_signal    2183..2238
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2599..2615)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2623..2639
                     /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(2647..2677)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     2652..2673
                     /label=lac-pro-F
     protein_bind    2692..2713
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2772..2967
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        2797..2967
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2818..2953
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2973..3107
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3346..3934)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3350..3369)
                     /label=pUBC-primer-1
     CDS             complement(4105..4965)
                     /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(4136..4157)
                     /label=Amp-F
     primer_bind     4879..4899
                     /label=Amp-R-seq
     promoter        complement(4966..5069)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5142..6354
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(5269..5291)
                     /label=hUBC-pro-R
     primer_bind     5737..5760
                     /label=pUBC-seq-2
     primer_bind     6191..6210
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
ORIGIN
        1 gaagcttggg ctgcaggtcg agctcaagct tcgaattctg cagtcgacgg taccgcgggc
       61 ccgggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
      121 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
      181 gggcgagggc gatgccacct acggcaagct gaccctgaag ctcatctgca ccaccggcaa
      241 gctgcccgtg ccctggccca ccctcgtgac caccctcggc tacggcctgc agtgcttcgc
      301 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
      361 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
      421 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
      481 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
      541 caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga
      601 ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
      661 cgtgctgctg cccgacaacc actacctgag ctaccagtcc aagctgagca aagaccccaa
      721 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
      781 catggacgag ctgtacaagt atccatacga cgtgcctgat tatgcaggcg gcagcggcgg
      841 caccggcggc agcggcggca ccatcgacta caccgccgcc ggcttcaccc tgctgcaggg
      901 cgcccacctg tacgcccccg aggacagggg catctgcgac gtgctggtgg ccaacggcaa
      961 gatcatcgcc gtggccagca acatccccag cgacatcgtg cccaactgca ccgtggtgga
     1021 cctgagcggc cagatcctgt gccccggctt catcgaccag cacgtgcacc tgatcggcgg
     1081 cggcggcgag gccggcccca ccaccaggac ccccgaggtg gccctgagca ggctgaccga
     1141 ggccggcgtg accagcgtgg tgggcctgct gggcaccgac agcatcagca ggcaccccga
     1201 gagcctgctg gccaagacca gggccctgaa cgaggagggc atcagcgcct ggatgctgac
     1261 cggcgcctac cacgtgccca gcaggaccat caccggcagc gtggagaagg acgtggccat
     1321 catcgacagg gtgatcggcg tgaagtgcgc catcagcgac cacaggagcg ccgcccccga
     1381 cgtgtaccac ctggccaaca tggccgccga gagcagggtg ggcggcctgc tgggcggcaa
     1441 gcccggcgtg accgtgttcc acatgggcga cagcaagaag gccctgcagc ccatctacga
     1501 cctgctggag aactgcgacg tgcccatcag caagctgctg cccacccacg tgaacaggaa
     1561 cgtgcccctg ttctaccagg ccctggagtt cgccaggaag ggcggcacca tcgacatcac
     1621 cagcagcatc gacgagcccg tggcccccgc cgagggcatc gccagggccg tgcaggccgg
     1681 catccccctg gccagggtga ccctgagcag cgacggcaac ggcagccagc ccttcttcga
     1741 cgacgagggc aacctgaccc acatcggcgt ggccggcttc gagaccctgc tggagaccgt
     1801 gcaggtgctg gtgaaggact acgacttcag catcagcgac gccctgaggc ccctgaccag
     1861 cagcgtggcc ggcttcctga acctgaccgg caagggcgag atcctgcccg gcaacgacgc
     1921 cgacctgctg gtgatgaccc ccgagctgag gatcgagcag gtgtacgcca ggggcaagct
     1981 gatggtgaag gacggcaagg cctgcgtgaa gggcaccttc gagacggcct gagcggccgc
     2041 actcctcagg tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct
     2101 cacaaatacc actgagatct ttttccctct gccaaaaatt atggggacat catgaagccc
     2161 cttgagcatc tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg
     2221 aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag
     2281 aatgagtatt tggtttagag tttggcaaca tatgcccata tgctggctgc catgaacaaa
     2341 ggttggctat aaagaggtca tcagtatatg aaacagcccc ctgctgtcca ttccttattc
     2401 catagaaaag ccttgacttg aggttagatt ttttttatat tttgttttgt gttatttttt
     2461 tctttaacat ccctaaaatt ttccttacat gttttactag ccagattttt cctcctctcc
     2521 tgactactcc cagtcatagc tgtccctctt ctcttatgga gatccctcga cctgcagccc
     2581 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     2641 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     2701 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     2761 ccagcggatc cgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     2821 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     2881 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     2941 ttttttggag gcctaggctt ttgcaaaaag ctaacttgtt tattgcagct tataatggtt
     3001 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3061 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatccgc tgcattaatg
     3121 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     3181 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3241 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3301 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3361 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3421 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3481 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3541 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3601 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3661 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3721 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3781 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3841 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3901 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3961 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4021 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4081 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4141 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4201 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4261 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4321 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4381 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4441 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4501 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4561 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4621 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4681 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4741 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4801 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4861 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4921 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4981 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5041 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgggtcg
     5101 acattgatta ttgactagta acccgtgtcg gctccagatc tggcctccgc gccgggtttt
     5161 ggcgcctccc gcgggcgccc ccctcctcac ggcgagcgct gccacgtcag acgaagggcg
     5221 cagcgagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga
     5281 ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc
     5341 tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg
     5401 cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc
     5461 gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga
     5521 tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc
     5581 gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt
     5641 ccgcgagcaa ggttgccctg aactgggggt tgggggggag cgcagcaaaa tggcggctgt
     5701 tcccgagtct tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg aaacaaggtg
     5761 gggggcatgg tgggcggcaa gaacccaagg tcttgaggcc ttcgctaatg cgggaaagct
     5821 cttattcggg tgagatgggc tggggcacca tctggggacc ctgacgtgaa gtttgtcact
     5881 gactggagaa ctcggtttgt cgtctgttgc gggggcggca gttatggcgg tgccgttggg
     5941 cagtgcaccc gtacctttgg gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg
     6001 ttggcttata atgcagggtg gggccacctg ccggtaggtg tgcggtaggc ttttctccgt
     6061 cgcaggacgc agggttcggg cctagggtag gctctcctga atcgacaggc gccggacctc
     6121 tggtgagggg agggataagt gaggcgtcag tttctttggt cggttttatg tacctatctt
     6181 cttaagtagc tgaagctccg gttttgaact atgcgctcgg ggttggcgag tgtgttttgt
     6241 gaagtttttt aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt
     6301 tagactagta aattgtccgc taaattctgg ccgtttttgg cttttttgtt agac
//