pUBC-2WCV-EGFP

LOCUS       pUBC-2WCV-EGFP        5550 bp DNA     circular SYN 11-APR-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5550)
  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..5550
                     /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..480
                     /codon_start=1
                     /label=2WCV(E77Y)
                     /translation="MLKTISPLISPELLKVLAEMGHGDEIIFSDAHFPAHSMGPQVIRA
                     DGLLVSDLLQAIIPLFELDSYAPPLVMMAAVYGDTLDPEVERRYRNALSLQAPCPDIIR
                     INRFAFYERAQKAFAIVITGERAKYGNILLKKGVTP"
     CDS             481..510
                     /codon_start=1
                     /label=Linker10
                     /translation="GGGGSGGGGS"
     CDS             512..1227
                     /codon_start=1
                     /label=EGFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     polyA_signal    1379..1434
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1795..1811)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1819..1835
                     /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(1843..1873)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     1848..1869
                     /label=lac-pro-F
     protein_bind    1888..1909
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1968..2163
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        1993..2163
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2014..2149
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2169..2303
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2542..3130)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2546..2565)
                     /label=pUBC-primer-1
     CDS             complement(3301..4161)
                     /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(3332..3353)
                     /label=Amp-F
     primer_bind     4075..4095
                     /label=Amp-R-seq
     promoter        complement(4162..4265)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4338..4737
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(4465..4487)
                     /label=hUBC-pro-R
     primer_bind     4933..4956
                     /label=pUBC-seq-2
     promoter        5024..5550
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5387..5406
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
ORIGIN
        1 gaagcttggg ctgcaggtcg agctcaagct tcgaattctg cagtcgacgg taccgccacc
       61 atgctgaaga ccatcagccc cctgatcagc cccgagctgc tgaaggtgct ggccgagatg
      121 ggccacggcg acgagatcat cttcagcgac gcccacttcc ccgcccacag catgggcccc
      181 caggtgatca gggccgacgg cctgctggtg agcgacctgc tgcaggccat catccccctg
      241 ttcgagctgg acagctacgc cccccccctg gtgatgatgg ccgccgtgta cggcgacacc
      301 ctggaccccg aggtggagag gaggtacagg aacgccctga gcctgcaggc cccctgcccc
      361 gacatcatca ggatcaacag gttcgccttc tacgagaggg cccagaaggc cttcgccatc
      421 gtgatcaccg gcgagagggc caagtacggc aacatcctgc tgaagaaggg cgtgaccccc
      481 ggcggcggcg gcagcggcgg cggcggcagc gtgagcaagg gcgaggagct gttcaccggg
      541 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
      601 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
      661 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
      721 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
      781 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
      841 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
      901 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
      961 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     1021 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     1081 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     1141 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     1201 ctcggcatgg acgagctgta caagtaaagc ggccgcactc ctcaggtgca ggctgcctat
     1261 cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt
     1321 ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa
     1381 taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga
     1441 aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg
     1501 gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag
     1561 tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt
     1621 tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc
     1681 ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc
     1741 cctcttctct tatggagatc cctcgacctg cagcccaagc ttggcgtaat catggtcata
     1801 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     1861 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     1921 ctcactgccc gctttccagt cgggaaacct gtcgtgccag cggatccgca tctcaattag
     1981 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     2041 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     2101 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     2161 aaaaagctaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     2221 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     2281 atgtatctta tcatgtctgg atccgctgca ttaatgaatc ggccaacgcg cggggagagg
     2341 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2401 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2461 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2521 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2581 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2641 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2701 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     2761 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2821 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     2881 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2941 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3001 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3061 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3121 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3181 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3241 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3301 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3361 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3421 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3481 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3541 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3601 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3661 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     3721 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     3781 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     3841 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     3901 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     3961 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4021 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4081 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4141 acggaaatgt tgaatactca tactcttcct ttttcatatt attgaagcat ttatcagggt
     4201 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4261 ccgcgcacat ttccccgaaa agtgccacct gggtcgacat tgattattga ctagtaaccc
     4321 gtgtcggctc cagatctggc ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct
     4381 cctcacggcg agcgctgcca cgtcagacga agggcgcagc gagcgtcctg atccttccgc
     4441 ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca
     4501 gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt tctttccaga
     4561 gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg
     4621 gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc
     4681 gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag
     4741 tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa
     4801 gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact
     4861 gggggttggg ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc
     4921 ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac
     4981 ccaaggtctt gaggccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg
     5041 gcaccatctg gggaccctga cgtgaagttt gtcactgact ggagaactcg gtttgtcgtc
     5101 tgttgcgggg gcggcagtta tggcggtgcc gttgggcagt gcacccgtac ctttgggagc
     5161 gcgcgccctc gtcgtgtcgt gacgtcaccc gttctgttgg cttataatgc agggtggggc
     5221 cacctgccgg taggtgtgcg gtaggctttt ctccgtcgca ggacgcaggg ttcgggccta
     5281 gggtaggctc tcctgaatcg acaggcgccg gacctctggt gaggggaggg ataagtgagg
     5341 cgtcagtttc tttggtcggt tttatgtacc tatcttctta agtagctgaa gctccggttt
     5401 tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc accttttgaa
     5461 atgtaatcat ttgggtcaat atgtaatttt cagtgttaga ctagtaaatt gtccgctaaa
     5521 ttctggccgt ttttggcttt tttgttagac
//