pUBC-mTagBFP2-1POK

LOCUS       pUBC-mTagBFP2-1POK            6351 bp DNA     circular SYN 21-MAR-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6351)
  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..6351
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     57..100
                     /label=Primer 1
     regulatory      77..86
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             83..781
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(749..823)
                     /label=mTagBFP2-V5-R
     CDS             782..823
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             824..859
                     /codon_start=1
                     /label=Linker12
                     /translation="GGSGGTGGSGGT"
     CDS             860..2026
                     /codon_start=1
                     /label=1P0K(E239Y)
                     /translation="IDYTAAGFTLLQGAHLYAPEDRGICDVLVANGKIIAVASNIPSDI
                     VPNCTVVDLSGQILCPGFIDQHVHLIGGGGEAGPTTRTPEVALSRLTEAGVTSVVGLLG
                     TDSISRHPESLLAKTRALNEEGISAWMLTGAYHVPSRTITGSVEKDVAIIDRVIGVKCA
                     ISDHRSAAPDVYHLANMAAESRVGGLLGGKPGVTVFHMGDSKKALQPIYDLLENCDVPI
                     SKLLPTHVNRNVPLFYQALEFARKGGTIDITSSIDEPVAPAEGIARAVQAGIPLARVTL
                     SSDGNGSQPFFDDEGNLTHIGVAGFETLLETVQVLVKDYDFSISDALRPLTSSVAGFLN
                     LTGKGEILPGNDADLLVMTPELRIEQVYARGKLMVKDGKACVKGTFETA"
     primer_bind     complement(2003..2050)
                     /label=NOtI-1POK-R
     polyA_signal    2180..2235
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2596..2612)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2620..2636
                     /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(2644..2674)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     2649..2670
                     /label=lac-pro-F
     protein_bind    2689..2710
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2769..2964
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        2794..2964
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2815..2950
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2970..3104
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3343..3931)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3347..3366)
                     /label=pUBC-primer-1
     CDS             complement(4102..4962)
                     /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(4133..4154)
                     /label=Amp-F
     primer_bind     4876..4896
                     /label=Amp-R-seq
     promoter        complement(4963..5066)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5139..5538
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(5266..5288)
                     /label=hUBC-pro-R
     primer_bind     5734..5757
                     /label=pUBC-seq-2
     promoter        5825..6351
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6188..6207
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
ORIGIN
        1 gaagcttggg ctgcaggtcg agctcaagct tcgaattctg cagtcgacgg taccgcgggc
       61 ccgggatcca ccggtcgcca ccatgagcga gctgattaag gagaacatgc acatgaagct
      121 gtacatggag ggcaccgtgg acaaccatca cttcaagtgc acatccgagg gcgaaggcaa
      181 gccctacgag ggcacccaga ccatgagaat caaggtggtc gagggcggcc ctctcccctt
      241 cgccttcgac atcctggcta ctagcttcct ctacggcagc aagaccttca tcaaccacac
      301 ccagggcatc cccgacttct tcaagcagtc cttccctgag ggcttcacat gggagagagt
      361 caccacatac gaagacgggg gcgtgctgac cgctacccag gacaccagcc tccaggacgg
      421 ctgcctcatc tacaacgtca agatcagagg ggtgaacttc acatccaacg gccctgtgat
      481 gcagaagaaa acactcggct gggaggcctt caccgagacg ctgtaccccg ctgacggcgg
      541 cctggaaggc agaaacgaca tggccctgaa gctcgtgggc gggagccatc tgatcgcaaa
      601 cgccaagacc acatatagat ccaagaaacc cgctaagaac ctcaagatgc ctggcgtcta
      661 ctatgtggac tacagactgg aaagaatcaa ggaggccaac aacgagacct acgtcgagca
      721 gcacgaggtg gcagtggcca gatactgcga cctccctagc aaactggggc acaagcttaa
      781 tggtaagcct atccctaacc ctctcctcgg tctcgattct acgggcggca gcggcggcac
      841 cggcggcagc ggcggcacca tcgactacac cgccgccggc ttcaccctgc tgcagggcgc
      901 ccacctgtac gcccccgagg acaggggcat ctgcgacgtg ctggtggcca acggcaagat
      961 catcgccgtg gccagcaaca tccccagcga catcgtgccc aactgcaccg tggtggacct
     1021 gagcggccag atcctgtgcc ccggcttcat cgaccagcac gtgcacctga tcggcggcgg
     1081 cggcgaggcc ggccccacca ccaggacccc cgaggtggcc ctgagcaggc tgaccgaggc
     1141 cggcgtgacc agcgtggtgg gcctgctggg caccgacagc atcagcaggc accccgagag
     1201 cctgctggcc aagaccaggg ccctgaacga ggagggcatc agcgcctgga tgctgaccgg
     1261 cgcctaccac gtgcccagca ggaccatcac cggcagcgtg gagaaggacg tggccatcat
     1321 cgacagggtg atcggcgtga agtgcgccat cagcgaccac aggagcgccg cccccgacgt
     1381 gtaccacctg gccaacatgg ccgccgagag cagggtgggc ggcctgctgg gcggcaagcc
     1441 cggcgtgacc gtgttccaca tgggcgacag caagaaggcc ctgcagccca tctacgacct
     1501 gctggagaac tgcgacgtgc ccatcagcaa gctgctgccc acccacgtga acaggaacgt
     1561 gcccctgttc taccaggccc tggagttcgc caggaagggc ggcaccatcg acatcaccag
     1621 cagcatcgac gagcccgtgg cccccgccga gggcatcgcc agggccgtgc aggccggcat
     1681 ccccctggcc agggtgaccc tgagcagcga cggcaacggc agccagccct tcttcgacga
     1741 cgagggcaac ctgacccaca tcggcgtggc cggcttcgag accctgctgg agaccgtgca
     1801 ggtgctggtg aaggactacg acttcagcat cagcgacgcc ctgaggcccc tgaccagcag
     1861 cgtggccggc ttcctgaacc tgaccggcaa gggcgagatc ctgcccggca acgacgccga
     1921 cctgctggtg atgacccccg agctgaggat cgagcaggtg tacgccaggg gcaagctgat
     1981 ggtgaaggac ggcaaggcct gcgtgaaggg caccttcgag acggcctgag cggccgcact
     2041 cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac
     2101 aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt
     2161 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     2221 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat
     2281 gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt
     2341 tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat
     2401 agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct
     2461 ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga
     2521 ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacct gcagcccaag
     2581 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     2641 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     2701 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     2761 gcggatccgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     2821 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     2881 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     2941 tttggaggcc taggcttttg caaaaagcta acttgtttat tgcagcttat aatggttaca
     3001 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     3061 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatccgctgc attaatgaat
     3121 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3181 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3241 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3301 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3361 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3421 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3481 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3541 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3601 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3661 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3721 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3781 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3841 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3901 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3961 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4021 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4081 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4141 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4201 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4261 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4321 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4381 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4441 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4501 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4561 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4621 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4681 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4741 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4801 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4861 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4921 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcatat
     4981 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5041 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgggtcgaca
     5101 ttgattattg actagtaacc cgtgtcggct ccagatctgg cctccgcgcc gggttttggc
     5161 gcctcccgcg ggcgcccccc tcctcacggc gagcgctgcc acgtcagacg aagggcgcag
     5221 cgagcgtcct gatccttccg cccggacgct caggacagcg gcccgctgct cataagactc
     5281 ggccttagaa ccccagtatc agcagaagga cattttagga cgggacttgg gtgactctag
     5341 ggcactggtt ttctttccag agagcggaac aggcgaggaa aagtagtccc ttctcggcga
     5401 ttctgcggag ggatctccgt ggggcggtga acgccgatga ttatataagg acgcgccggg
     5461 tgtggcacag ctagttccgt cgcagccggg atttgggtcg cggttcttgt ttgtggatcg
     5521 ctgtgatcgt cacttggtga gtagcgggct gctgggctgg ccggggcttt cgtggccgcc
     5581 gggccgctcg gtgggacgga agcgtgtgga gagaccgcca agggctgtag tctgggtccg
     5641 cgagcaaggt tgccctgaac tgggggttgg gggggagcgc agcaaaatgg cggctgttcc
     5701 cgagtcttga atggaagacg cttgtgaggc gggctgtgag gtcgttgaaa caaggtgggg
     5761 ggcatggtgg gcggcaagaa cccaaggtct tgaggccttc gctaatgcgg gaaagctctt
     5821 attcgggtga gatgggctgg ggcaccatct ggggaccctg acgtgaagtt tgtcactgac
     5881 tggagaactc ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag
     5941 tgcacccgta cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg
     6001 gcttataatg cagggtgggg ccacctgccg gtaggtgtgc ggtaggcttt tctccgtcgc
     6061 aggacgcagg gttcgggcct agggtaggct ctcctgaatc gacaggcgcc ggacctctgg
     6121 tgaggggagg gataagtgag gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt
     6181 aagtagctga agctccggtt ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa
     6241 gttttttagg caccttttga aatgtaatca tttgggtcaa tatgtaattt tcagtgttag
     6301 actagtaaat tgtccgctaa attctggccg tttttggctt ttttgttaga c
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