pUBC-mEGFP-1POK

LOCUS       pUBC-mEGFP-1POK               6375 bp DNA     circular SYN 25-APR-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6375)
  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..6375
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      55..60
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             61..1230
                     /codon_start=1
                     /label=1P0K(E239Y)
                     /translation="MIDYTAAGFTLLQGAHLYAPEDRGICDVLVANGKIIAVASNIPSD
                     IVPNCTVVDLSGQILCPGFIDQHVHLIGGGGEAGPTTRTPEVALSRLTEAGVTSVVGLL
                     GTDSISRHPESLLAKTRALNEEGISAWMLTGAYHVPSRTITGSVEKDVAIIDRVIGVKC
                     AISDHRSAAPDVYHLANMAAESRVGGLLGGKPGVTVFHMGDSKKALQPIYDLLENCDVP
                     ISKLLPTHVNRNVPLFYQALEFARKGGTIDITSSIDEPVAPAEGIARAVQAGIPLARVT
                     LSSDGNGSQPFFDDEGNLTHIGVAGFETLLETVQVLVKDYDFSISDALRPLTSSVAGFL
                     NLTGKGEILPGNDADLLVMTPELRIEQVYARGKLMVKDGKACVKGTFETA"
     transit_peptide 1231..1305
                     /codon_start=1
                     /label=Linker25
                     /translation="GGSGGTGGSGGTGGSGGTGGSGGTG"
     primer_bind     1286..1353
                     /label=Primer 1
     CDS             1306..1332
                     /codon_start=1
                     /label=HA tag
                     /translation="YPYDVPDYA"
     CDS             1336..2049
                     /codon_start=1
                     /label=mEGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1935..1969)
                     /label=mEGFP-Qch-R
     primer_bind     1935..1969
                     /label=mEGFP-Qch-F
     misc_feature    1951..1953
                     /label=A to K for mEGFP
     primer_bind     complement(2031..2074)
                     /label=mEGFP-R
     polyA_signal    2204..2259
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2620..2636)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2644..2660
                     /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(2668..2698)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     2673..2694
                     /label=lac-pro-F
     protein_bind    2713..2734
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2793..2988
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        2818..2988
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2839..2974
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2994..3128
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3367..3955)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3371..3390)
                     /label=pUBC-primer-1
     CDS             complement(4126..4986)
                     /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(4157..4178)
                     /label=Amp-F
     primer_bind     4900..4920
                     /label=Amp-R-seq
     promoter        complement(4987..5090)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5163..5562
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(5290..5312)
                     /label=hUBC-pro-R
     primer_bind     5758..5781
                     /label=pUBC-seq-2
     promoter        5849..6375
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6212..6231
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
ORIGIN
        1 gaagcttggg ctgcaggtcg agctcaagct tcgaattctg cagtcgacgg taccgccacc
       61 atgatcgact acaccgccgc cggcttcacc ctgctgcagg gcgcccacct gtacgccccc
      121 gaggacaggg gcatctgcga cgtgctggtg gccaacggca agatcatcgc cgtggccagc
      181 aacatcccca gcgacatcgt gcccaactgc accgtggtgg acctgagcgg ccagatcctg
      241 tgccccggct tcatcgacca gcacgtgcac ctgatcggcg gcggcggcga ggccggcccc
      301 accaccagga cccccgaggt ggccctgagc aggctgaccg aggccggcgt gaccagcgtg
      361 gtgggcctgc tgggcaccga cagcatcagc aggcaccccg agagcctgct ggccaagacc
      421 agggccctga acgaggaggg catcagcgcc tggatgctga ccggcgccta ccacgtgccc
      481 agcaggacca tcaccggcag cgtggagaag gacgtggcca tcatcgacag ggtgatcggc
      541 gtgaagtgcg ccatcagcga ccacaggagc gccgcccccg acgtgtacca cctggccaac
      601 atggccgccg agagcagggt gggcggcctg ctgggcggca agcccggcgt gaccgtgttc
      661 cacatgggcg acagcaagaa ggccctgcag cccatctacg acctgctgga gaactgcgac
      721 gtgcccatca gcaagctgct gcccacccac gtgaacagga acgtgcccct gttctaccag
      781 gccctggagt tcgccaggaa gggcggcacc atcgacatca ccagcagcat cgacgagccc
      841 gtggcccccg ccgagggcat cgccagggcc gtgcaggccg gcatccccct ggccagggtg
      901 accctgagca gcgacggcaa cggcagccag cccttcttcg acgacgaggg caacctgacc
      961 cacatcggcg tggccggctt cgagaccctg ctggagaccg tgcaggtgct ggtgaaggac
     1021 tacgacttca gcatcagcga cgccctgagg cccctgacca gcagcgtggc cggcttcctg
     1081 aacctgaccg gcaagggcga gatcctgccc ggcaacgacg ccgacctgct ggtgatgacc
     1141 cccgagctga ggatcgagca ggtgtacgcc aggggcaagc tgatggtgaa ggacggcaag
     1201 gcctgcgtga agggcacctt cgagacggcc ggcggaagcg gcgggaccgg cggcagcgga
     1261 gggaccgggg ggagcggcgg caccggagga agcgggggaa ccggttatcc atacgacgtg
     1321 cctgattatg caggcgtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     1381 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     1441 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     1501 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     1561 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     1621 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     1681 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1741 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1801 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1861 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1921 cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc
     1981 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     2041 ctgtacaagt gaagcggccg cactcctcag gtgcaggctg cctatcagaa ggtggtggct
     2101 ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat
     2161 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
     2221 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
     2281 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat
     2341 atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc
     2401 cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata
     2461 ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta
     2521 gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg
     2581 agatccctcg acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     2641 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     2701 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     2761 ccagtcggga aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc
     2821 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     2881 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     2941 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt
     3001 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3061 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     3121 tctggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3181 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3241 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3301 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3361 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3421 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3481 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3541 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3601 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3661 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3721 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3781 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3841 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3901 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3961 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4021 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4081 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4141 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4201 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4261 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4321 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4381 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4441 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4501 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4561 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4621 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4681 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4741 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4801 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4861 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4921 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4981 actcatactc ttcctttttc atattattga agcatttatc agggttattg tctcatgagc
     5041 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     5101 cgaaaagtgc cacctgggtc gacattgatt attgactagt aacccgtgtc ggctccagat
     5161 ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcctca cggcgagcgc
     5221 tgccacgtca gacgaagggc gcagcgagcg tcctgatcct tccgcccgga cgctcaggac
     5281 agcggcccgc tgctcataag actcggcctt agaaccccag tatcagcaga aggacatttt
     5341 aggacgggac ttgggtgact ctagggcact ggttttcttt ccagagagcg gaacaggcga
     5401 ggaaaagtag tcccttctcg gcgattctgc ggagggatct ccgtggggcg gtgaacgccg
     5461 atgattatat aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg
     5521 gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg
     5581 ctggccgggg ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg tggagagacc
     5641 gccaagggct gtagtctggg tccgcgagca aggttgccct gaactggggg ttggggggga
     5701 gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa gacgcttgtg aggcgggctg
     5761 tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca agaacccaag gtcttgaggc
     5821 cttcgctaat gcgggaaagc tcttattcgg gtgagatggg ctggggcacc atctggggac
     5881 cctgacgtga agtttgtcac tgactggaga actcggtttg tcgtctgttg cgggggcggc
     5941 agttatggcg gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg ccctcgtcgt
     6001 gtcgtgacgt cacccgttct gttggcttat aatgcagggt ggggccacct gccggtaggt
     6061 gtgcggtagg cttttctccg tcgcaggacg cagggttcgg gcctagggta ggctctcctg
     6121 aatcgacagg cgccggacct ctggtgaggg gagggataag tgaggcgtca gtttctttgg
     6181 tcggttttat gtacctatct tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg
     6241 gggttggcga gtgtgttttg tgaagttttt taggcacctt ttgaaatgta atcatttggg
     6301 tcaatatgta attttcagtg ttagactagt aaattgtccg ctaaattctg gccgtttttg
     6361 gcttttttgt tagac
//