pUBC-EBFP2-1POK

LOCUS       pUBC-EBFP2-1POK              6369 bp DNA     circular SYN 26-APR-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6369)
  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..6369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     55..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..799
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(128..149)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(770..841)
                     /label=EBFP2-V5-R
     primer_bind     800..862
                     /label=V5-1POK-F
     CDS             800..841
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             842..877
                     /codon_start=1
                     /label=Linker12
                     /translation="GGSGGTGGSGGT"
     CDS             878..2044
                     /codon_start=1
                     /label=1P0K(E239Y)
                     /translation="IDYTAAGFTLLQGAHLYAPEDRGICDVLVANGKIIAVASNIPSDI
                     VPNCTVVDLSGQILCPGFIDQHVHLIGGGGEAGPTTRTPEVALSRLTEAGVTSVVGLLG
                     TDSISRHPESLLAKTRALNEEGISAWMLTGAYHVPSRTITGSVEKDVAIIDRVIGVKCA
                     ISDHRSAAPDVYHLANMAAESRVGGLLGGKPGVTVFHMGDSKKALQPIYDLLENCDVPI
                     SKLLPTHVNRNVPLFYQALEFARKGGTIDITSSIDEPVAPAEGIARAVQAGIPLARVTL
                     SSDGNGSQPFFDDEGNLTHIGVAGFETLLETVQVLVKDYDFSISDALRPLTSSVAGFLN
                     LTGKGEILPGNDADLLVMTPELRIEQVYARGKLMVKDGKACVKGTFETA"
     primer_bind     1722..1738
                     /label=V5-1POK-F
     primer_bind     complement(2021..2068)
                     /label=NOtI-1POK-R
     polyA_signal    2198..2253
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2614..2630)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2638..2654
                     /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(2662..2692)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     2667..2688
                     /label=lac-pro-F
     protein_bind    2707..2728
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2787..2982
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        2812..2982
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2833..2968
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2988..3122
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3361..3949)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3365..3384)
                     /label=pUBC-primer-1
     CDS             complement(4120..4980)
                     /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(4151..4172)
                     /label=Amp-F
     primer_bind     4894..4914
                     /label=Amp-R-seq
     promoter        complement(4981..5084)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5157..5556
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(5284..5306)
                     /label=hUBC-pro-R
     primer_bind     5752..5775
                     /label=pUBC-seq-2
     promoter        5843..6369
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6206..6225
                     /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 tgaggggcga
      181 gggcgagggc gatgccacca acggcaagct gaccctgaag ttcatctgca ccaccggcaa
      241 gctgcccgtg ccctggccca ccctcgtgac caccctgagc cacggcgtgc agtgcttcgc
      301 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
      361 cgtccaggag cgcaccatct tcttcaagga cgacggcacc tacaagaccc gcgccgaggt
      421 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcgtcg acttcaagga
      481 ggacggcaac atcctggggc acaagctgga gtacaacttc aacagccaca acatctatat
      541 catggccgtc aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacgtgga
      601 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
      661 cgtgctgctg cccgacagcc actacctgag cacccagtcc gtgctgagca aagaccccaa
      721 cgagaagcgc gatcacatgg tcctgctgga gttccgcacc gccgccggga tcactctcgg
      781 catggacgag ctgtacaagg gtaagcctat ccctaaccct ctcctcggtc tcgattctac
      841 gggcggcagc ggcggcaccg gcggcagcgg cggcaccatc gactacaccg ccgccggctt
      901 caccctgctg cagggcgccc acctgtacgc ccccgaggac aggggcatct gcgacgtgct
      961 ggtggccaac ggcaagatca tcgccgtggc cagcaacatc cccagcgaca tcgtgcccaa
     1021 ctgcaccgtg gtggacctga gcggccagat cctgtgcccc ggcttcatcg accagcacgt
     1081 gcacctgatc ggcggcggcg gcgaggccgg ccccaccacc aggacccccg aggtggccct
     1141 gagcaggctg accgaggccg gcgtgaccag cgtggtgggc ctgctgggca ccgacagcat
     1201 cagcaggcac cccgagagcc tgctggccaa gaccagggcc ctgaacgagg agggcatcag
     1261 cgcctggatg ctgaccggcg cctaccacgt gcccagcagg accatcaccg gcagcgtgga
     1321 gaaggacgtg gccatcatcg acagggtgat cggcgtgaag tgcgccatca gcgaccacag
     1381 gagcgccgcc cccgacgtgt accacctggc caacatggcc gccgagagca gggtgggcgg
     1441 cctgctgggc ggcaagcccg gcgtgaccgt gttccacatg ggcgacagca agaaggccct
     1501 gcagcccatc tacgacctgc tggagaactg cgacgtgccc atcagcaagc tgctgcccac
     1561 ccacgtgaac aggaacgtgc ccctgttcta ccaggccctg gagttcgcca ggaagggcgg
     1621 caccatcgac atcaccagca gcatcgacga gcccgtggcc cccgccgagg gcatcgccag
     1681 ggccgtgcag gccggcatcc ccctggccag ggtgaccctg agcagcgacg gcaacggcag
     1741 ccagcccttc ttcgacgacg agggcaacct gacccacatc ggcgtggccg gcttcgagac
     1801 cctgctggag accgtgcagg tgctggtgaa ggactacgac ttcagcatca gcgacgccct
     1861 gaggcccctg accagcagcg tggccggctt cctgaacctg accggcaagg gcgagatcct
     1921 gcccggcaac gacgccgacc tgctggtgat gacccccgag ctgaggatcg agcaggtgta
     1981 cgccaggggc aagctgatgg tgaaggacgg caaggcctgc gtgaagggca ccttcgagac
     2041 ggcctgagcg gccgcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg
     2101 gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg
     2161 gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt
     2221 gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat
     2281 catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg
     2341 gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct
     2401 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt
     2461 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga
     2521 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc
     2581 ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     2641 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     2701 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     2761 gggaaacctg tcgtgccagc ggatccgcat ctcaattagt cagcaaccat agtcccgccc
     2821 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     2881 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     2941 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctaac ttgtttattg
     3001 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3061 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     3121 tccgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3181 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3241 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3301 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3361 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3421 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3481 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3541 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3601 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3661 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3721 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3781 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3841 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3901 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3961 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4021 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4081 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4141 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4201 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4261 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4321 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4381 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4441 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4501 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4561 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4621 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4681 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4741 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4801 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4861 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4921 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4981 actcttcctt tttcatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5041 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     5101 gtgccacctg ggtcgacatt gattattgac tagtaacccg tgtcggctcc agatctggcc
     5161 tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc ctcacggcga gcgctgccac
     5221 gtcagacgaa gggcgcagcg agcgtcctga tccttccgcc cggacgctca ggacagcggc
     5281 ccgctgctca taagactcgg ccttagaacc ccagtatcag cagaaggaca ttttaggacg
     5341 ggacttgggt gactctaggg cactggtttt ctttccagag agcggaacag gcgaggaaaa
     5401 gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt
     5461 atataaggac gcgccgggtg tggcacagct agttccgtcg cagccgggat ttgggtcgcg
     5521 gttcttgttt gtggatcgct gtgatcgtca cttggtgagt agcgggctgc tgggctggcc
     5581 ggggctttcg tggccgccgg gccgctcggt gggacggaag cgtgtggaga gaccgccaag
     5641 ggctgtagtc tgggtccgcg agcaaggttg ccctgaactg ggggttgggg gggagcgcag
     5701 caaaatggcg gctgttcccg agtcttgaat ggaagacgct tgtgaggcgg gctgtgaggt
     5761 cgttgaaaca aggtgggggg catggtgggc ggcaagaacc caaggtcttg aggccttcgc
     5821 taatgcggga aagctcttat tcgggtgaga tgggctgggg caccatctgg ggaccctgac
     5881 gtgaagtttg tcactgactg gagaactcgg tttgtcgtct gttgcggggg cggcagttat
     5941 ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg tcgtgtcgtg
     6001 acgtcacccg ttctgttggc ttataatgca gggtggggcc acctgccggt aggtgtgcgg
     6061 taggcttttc tccgtcgcag gacgcagggt tcgggcctag ggtaggctct cctgaatcga
     6121 caggcgccgg acctctggtg aggggaggga taagtgaggc gtcagtttct ttggtcggtt
     6181 ttatgtacct atcttcttaa gtagctgaag ctccggtttt gaactatgcg ctcggggttg
     6241 gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt tgggtcaata
     6301 tgtaattttc agtgttagac tagtaaattg tccgctaaat tctggccgtt tttggctttt
     6361 ttgttagac
//