pUBC-1M3U-EGFP

LOCUS       pUBC-1M3U-EGFP              5922 bp DNA     circular SYN 03-MAR-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5922)
  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..5922
                     /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..852
                     /codon_start=1
                     /label=1M3U(E158I, D161T)
                     /translation="MKPTTISLLQKYKQEKKRFATITAYDYSFAKLFADEGLNVMLVGD
                     SLGMTVQGHDSTLPVTVADIAYHTAAVRRGAPNCLLLADLPFMAYATPEQAFENAATVM
                     RAGANMVKIEGGEWLVETVQMLTERAVPVCGHLGLTPQSVNIFGGYKVQGRGDIAGTQL
                     LSDALALEAAGAQLLVLECVPVELAKRITEALAIPVIGIGAGNVTDGQILVMHDAFGIT
                     GGHIPKFAKNFLAETGDIRAAVRQYMAEVESGVYPGEEHSFH"
     CDS             853..882
                     /codon_start=1
                     /label=Linker10
                     /translation="GGGGSGGGGS"
     CDS             883..1599
                     /codon_start=3
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     polyA_signal    1751..1806
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2167..2183)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2191..2207
                     /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(2215..2245)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     2220..2241
                     /label=lac-pro-F
     protein_bind    2260..2281
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2340..2535
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        2365..2535
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2386..2521
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2541..2675
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2914..3502)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2918..2937)
                     /label=pUBC-primer-1
     CDS             complement(3673..4533)
                     /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(3704..3725)
                     /label=Amp-F
     primer_bind     4447..4467
                     /label=Amp-R-seq
     promoter        complement(4534..4637)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4710..5109
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(4837..4859)
                     /label=hUBC-pro-R
     primer_bind     5305..5328
                     /label=pUBC-seq-2
     promoter        5396..5922
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5759..5778
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
ORIGIN
        1 gaagcttggg ctgcaggtcg agctcaagct tcgaattctg cagtcgacgg taccgccacc
       61 atgaagccca ccaccatcag cctgctgcag aagtacaagc aggagaagaa gaggttcgcc
      121 accatcaccg cctacgacta cagcttcgcc aagctgttcg ccgacgaggg cctgaacgtg
      181 atgctggtgg gcgacagcct gggcatgacc gtgcagggcc acgacagcac cctgcccgtg
      241 accgtggccg acatcgccta ccacaccgcc gccgtgagga ggggcgcccc caactgcctg
      301 ctgctggccg acctgccctt catggcctac gccacccccg agcaggcctt cgagaacgcc
      361 gccaccgtga tgagggccgg cgccaacatg gtgaagatcg agggcggcga gtggctggtg
      421 gagaccgtgc agatgctgac cgagagggcc gtgcccgtgt gcggccacct gggcctgacc
      481 ccccagagcg tgaacatctt cggcggctac aaggtgcagg gcaggggcga catcgccggc
      541 acccagctgc tgagcgacgc cctggccctg gaggccgccg gcgcccagct gctggtgctg
      601 gagtgcgtgc ccgtggagct ggccaagagg atcaccgagg ccctggccat ccccgtgatc
      661 ggcatcggcg ccggcaacgt gaccgacggc cagatcctgg tgatgcacga cgccttcggc
      721 atcaccggcg gccacatccc caagttcgcc aagaacttcc tggccgagac cggcgacatc
      781 agggccgccg tgaggcagta catggccgag gtggagagcg gcgtgtaccc cggcgaggag
      841 cacagcttcc acggcggcgg cggcagcggc ggcggcggca gcgtgagcaa gggcgaggag
      901 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
      961 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     1021 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     1081 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1141 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1201 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1261 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1321 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     1381 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     1441 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     1501 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1561 gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgcac tcctcaggtg
     1621 caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac
     1681 tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg
     1741 acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt
     1801 ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg
     1861 gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa
     1921 agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc
     1981 ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc
     2041 ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca
     2101 gtcatagctg tccctcttct cttatggaga tccctcgacc tgcagcccaa gcttggcgta
     2161 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     2221 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     2281 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg
     2341 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     2401 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     2461 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     2521 ctaggctttt gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca
     2581 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     2641 ccaaactcat caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg
     2701 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     2761 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2821 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2881 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2941 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3001 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3061 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3121 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3181 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3241 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3301 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3361 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3421 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3481 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3541 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3601 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3661 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3721 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3781 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     3841 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3901 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3961 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4021 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4081 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4141 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4201 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4261 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4321 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4381 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4441 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4501 aataagggcg acacggaaat gttgaatact catactcttc ctttttcata ttattgaagc
     4561 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4621 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgggtcgac attgattatt
     4681 gactagtaac ccgtgtcggc tccagatctg gcctccgcgc cgggttttgg cgcctcccgc
     4741 gggcgccccc ctcctcacgg cgagcgctgc cacgtcagac gaagggcgca gcgagcgtcc
     4801 tgatccttcc gcccggacgc tcaggacagc ggcccgctgc tcataagact cggccttaga
     4861 accccagtat cagcagaagg acattttagg acgggacttg ggtgactcta gggcactggt
     4921 tttctttcca gagagcggaa caggcgagga aaagtagtcc cttctcggcg attctgcgga
     4981 gggatctccg tggggcggtg aacgccgatg attatataag gacgcgccgg gtgtggcaca
     5041 gctagttccg tcgcagccgg gatttgggtc gcggttcttg tttgtggatc gctgtgatcg
     5101 tcacttggtg agtagcgggc tgctgggctg gccggggctt tcgtggccgc cgggccgctc
     5161 ggtgggacgg aagcgtgtgg agagaccgcc aagggctgta gtctgggtcc gcgagcaagg
     5221 ttgccctgaa ctgggggttg ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg
     5281 aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg
     5341 ggcggcaaga acccaaggtc ttgaggcctt cgctaatgcg ggaaagctct tattcgggtg
     5401 agatgggctg gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact
     5461 cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt
     5521 acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat
     5581 gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag
     5641 ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag
     5701 ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg
     5761 aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag
     5821 gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gactagtaaa
     5881 ttgtccgcta aattctggcc gtttttggct tttttgttag ac
//