pUBC-1POK-MBP

LOCUS       pUBC-1POK-MBP             6765 bp DNA     circular SYN 27-MAY-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6765)
  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..6765
                     /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"
     primer_bind     495..504
                     /label=GFP-F1
     transit_peptide 1231..1305
                     /codon_start=1
                     /label=Linker25
                     /translation="GGSGGTGGSGGTGGSGGTGGSGGTG"
     CDS             1306..1332
                     /codon_start=1
                     /label=HA tag
                     /translation="YPYDVPDYA"
     CDS             1342..2439
                     /codon_start=1
                     /label=MBP_tag
                     /note="8 rare Homo sapiens codons were removed."
                     /translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     polyA_signal    2594..2649
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3010..3026)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3034..3050
                     /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(3058..3088)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     3063..3084
                     /label=lac-pro-F
     protein_bind    3103..3124
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3183..3378
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3208..3378
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3229..3364
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3384..3518
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3757..4345)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3761..3780)
                     /label=pUBC-primer-1
     CDS             complement(4516..5376)
                     /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(4547..4568)
                     /label=Amp-F
     primer_bind     5290..5310
                     /label=Amp-R-seq
     promoter        complement(5377..5480)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5553..5952
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(5680..5702)
                     /label=hUBC-pro-R
     primer_bind     6008..6020
                     /label=GFP-F1
     primer_bind     6148..6171
                     /label=pUBC-seq-2
     promoter        6239..6765
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6602..6621
                     /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 caggcagcgg caaaatcgag gaggggaagc tggtgatctg gattaatggc
     1381 gacaagggct acaatggcct ggccgaagtg ggcaagaagt tcgagaagga caccggcatc
     1441 aaggtgaccg tggagcaccc cgacaagctg gaggaaaagt tcccccaggt ggccgccacc
     1501 ggcgatggcc ctgacatcat cttttgggct cacgatagat tcggcggcta cgcccagagc
     1561 ggcctgctgg ccgagatcac ccccgataaa gccttccagg ataaactgta tcctttcacc
     1621 tgggatgccg tgagatacaa cggcaagctg atcgcctacc ccatcgccgt ggaagccctg
     1681 agcctgatct acaataagga cctgctgccc aaccctccaa agacctggga ggaaatcccc
     1741 gccctggata aggagctgaa ggccaagggc aagagcgccc tgatgtttaa tctgcaggag
     1801 ccctacttca cctggcccct gattgccgcc gacggcggct acgccttcaa gtacgagaat
     1861 ggcaagtatg acatcaagga tgtgggcgtg gacaatgccg gagccaaagc cggcctgacc
     1921 tttctggtgg acctgatcaa gaacaagcac atgaacgccg acaccgacta cagcatcgcc
     1981 gaggccgcct tcaataaggg cgagacagcc atgaccatca atggcccctg ggcctggagc
     2041 aacatcgaca ccagcaaggt gaattatggc gtgaccgtgc tgcccacctt caagggccag
     2101 cccagcaagc ccttcgtggg cgtgctgagc gccggcatca atgccgccag ccccaacaag
     2161 gagctggcca aggagttcct ggaaaactac ctgctgaccg atgagggcct ggaagccgtg
     2221 aacaaggaca aacccctggg agccgtggcc ctgaagagct atgaggaaga gctggccaag
     2281 gaccccagaa tcgccgccac catggagaat gcccagaagg gcgagatcat gcctaatatc
     2341 ccccagatga gcgccttctg gtatgccgtg agaaccgccg tgatcaatgc cgccagcggc
     2401 agacagaccg tggatgaggc cctgaaggat gcccagacct gaagcggccg cactcctcag
     2461 gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac
     2521 cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat
     2581 ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg
     2641 tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat
     2701 ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta
     2761 taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa
     2821 gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca
     2881 tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc
     2941 ccagtcatag ctgtccctct tctcttatgg agatccctcg acctgcagcc caagcttggc
     3001 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3061 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     3121 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat
     3181 ccgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3241 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3301 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3361 ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa
     3421 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3481 tgtccaaact catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca
     3541 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3601 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3661 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3721 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3781 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3841 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3901 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3961 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4021 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4081 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4141 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4201 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4261 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4321 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4381 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4441 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4501 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4561 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4621 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4681 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4741 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4801 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4861 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4921 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4981 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5041 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5101 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5161 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5221 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5281 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5341 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc atattattga
     5401 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5461 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgggtc gacattgatt
     5521 attgactagt aacccgtgtc ggctccagat ctggcctccg cgccgggttt tggcgcctcc
     5581 cgcgggcgcc cccctcctca cggcgagcgc tgccacgtca gacgaagggc gcagcgagcg
     5641 tcctgatcct tccgcccgga cgctcaggac agcggcccgc tgctcataag actcggcctt
     5701 agaaccccag tatcagcaga aggacatttt aggacgggac ttgggtgact ctagggcact
     5761 ggttttcttt ccagagagcg gaacaggcga ggaaaagtag tcccttctcg gcgattctgc
     5821 ggagggatct ccgtggggcg gtgaacgccg atgattatat aaggacgcgc cgggtgtggc
     5881 acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga
     5941 tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg ctttcgtggc cgccgggccg
     6001 ctcggtggga cggaagcgtg tggagagacc gccaagggct gtagtctggg tccgcgagca
     6061 aggttgccct gaactggggg ttggggggga gcgcagcaaa atggcggctg ttcccgagtc
     6121 ttgaatggaa gacgcttgtg aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg
     6181 gtgggcggca agaacccaag gtcttgaggc cttcgctaat gcgggaaagc tcttattcgg
     6241 gtgagatggg ctggggcacc atctggggac cctgacgtga agtttgtcac tgactggaga
     6301 actcggtttg tcgtctgttg cgggggcggc agttatggcg gtgccgttgg gcagtgcacc
     6361 cgtacctttg ggagcgcgcg ccctcgtcgt gtcgtgacgt cacccgttct gttggcttat
     6421 aatgcagggt ggggccacct gccggtaggt gtgcggtagg cttttctccg tcgcaggacg
     6481 cagggttcgg gcctagggta ggctctcctg aatcgacagg cgccggacct ctggtgaggg
     6541 gagggataag tgaggcgtca gtttctttgg tcggttttat gtacctatct tcttaagtag
     6601 ctgaagctcc ggttttgaac tatgcgctcg gggttggcga gtgtgttttg tgaagttttt
     6661 taggcacctt ttgaaatgta atcatttggg tcaatatgta attttcagtg ttagactagt
     6721 aaattgtccg ctaaattctg gccgtttttg gcttttttgt tagac
//