LOCUS       sequence-10530-e        5296 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5296)
  AUTHORS   Jodoin J, Misiewicz M, Makhijani P, Giannopoulos PN, Hammond J, 
            Goodyer CG, LeBlanc AC
  TITLE     Loss of anti-Bax function in gerstmann-Straussler-Scheinker 
            syndrome-associated prion protein mutants.
  JOURNAL   PLoS One. 2009  . 4(8):e6647.
  PUBMED    19680558
REFERENCE   2  (bases 1 to 5296)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One. 
            2009  . 4(8):e6647."
COMMENT     Sequence Label: pBud-EGFP
FEATURES             Location/Qualifiers
     source          1..5296
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        10..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     544..564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     638..657
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        638..656
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             719..748
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             764..781
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    812..933
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(823..842)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     877..896
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1063..1437)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(1456..1503)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(1551..1867)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1565..1700
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1619..1638)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1889..3051
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2120..3042
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2999..3019
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             join(3082..3084,3085..3087,3088..3801)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3082..3084,3085..3087,3088..3798)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3127..3148)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3388..3407)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3735..3756
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3828..3869
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3879..3896
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3919..3936)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3925..4149
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(4281..4869)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4361..4380)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4960..4978
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5016..5038)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5138..5157
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcgcgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
       61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      121 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      181 caatagggac tttccattga cgtcaatggg tggactattt acggtaaact gcccacttgg
      241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
      601 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
      661 cccaagcttg cattcctgca ggtcgacatc gatcttaagc agtacttcta gaggatccga
      721 acaaaaactc atctcagaag aggatctgaa tatgcatacc ggtcatcatc accatcacca
      781 ttgagtttga tccccgggaa ttcagacatg ataagataca ttgatgagtt tggacaaacc
      841 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
      901 tttgtaacca ttataagctg caataaacaa gttggggtgg gcgaagaact ccagcatgag
      961 atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct
     1021 ttcatagaag gcggcggtgg aatcgaaatc tcgtagcacg tgtcagtcct gctcctcggc
     1081 cacgaagtgc acgcagttgc cggccgggtc gcgcagggcg aactcccgcc cccacggctg
     1141 ctcgccgatc tcggtcatgg ccggcccgga ggcgtcccgg aagttcgtgg acacgacctc
     1201 cgaccactcg gcgtacagct cgtccaggcc gcgcacccac acccaggcca gggtgttgtc
     1261 cggcaccacc tggtcctgga ccgcgctgat gaacagggtc acgtcgtccc ggaccacacc
     1321 ggcgaagtcg tcctccacga agtcccggga gaacccgagc cggtcggtcc agaactcgac
     1381 cgctccggcg acgtcgcgcg cggtgagcac cggaacggca ctggtcaact tggccatggt
     1441 ttagttcctc accttgtcgt attatactat gccgatatac tatgccgatg attaattgtc
     1501 aacacgtgct gatcagatcc gaaaatggat atacaagctc ccgggagctt tttgcaaaag
     1561 cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc agaggcggcc
     1621 tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg
     1681 ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg
     1741 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt
     1801 gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt
     1861 ccacaccctc gtcgagctag cttcgtgagg ctccggtgcc cgtcagtggg cagagcgcac
     1921 atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaaccg gtgcctagag
     1981 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga
     2041 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg
     2101 gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg gcctctttac
     2161 gggttatggc ccttgcgtgc cttgaattac ttccacctgg ctccagtacg tgattcttga
     2221 tcccgagctg gagccagggg cgggccttgc gctttaggag ccccttcgcc tcgtgcttga
     2281 gttgaggcct ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc
     2341 tgtctcgctg ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg
     2401 ctttttttct ggcaagatag tcttgtaaat gcgggccagg atctgcacac tggtatttcg
     2461 gtttttgggc ccgcggccgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg
     2521 cggggcctgc gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct
     2581 gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc
     2641 cggtcggcac cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tccagggggc
     2701 tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa
     2761 agggcctttc cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc
     2821 aggcacctcg attagttctg gagcttttgg agtacgtcgt ctttaggttg gggggagggg
     2881 ttttatgcga tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg
     2941 cacttgatgt aattctcgtt ggaatttgcc ctttttgagt ttggatcttg gttcattctc
     3001 aagcctcaga cagtggttca aagttttttt cttccatttc aggtgtcgtg aacacgtggt
     3061 cgcggccgct tcgaaggtac catggtgagc aagggcgagg agctgttcac cggggtggtg
     3121 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     3181 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     3241 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     3301 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     3361 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     3421 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     3481 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     3541 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     3601 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     3661 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     3721 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     3781 atggacgagc tgtacaagta gatctggccg gctgggcccg tttcgaaggt aagcctatcc
     3841 ctaaccctct cctcggtctc gattctacgc gtaccggtca tcatcaccat caccattgag
     3901 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     3961 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     4021 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     4081 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     4141 gctctatggc ttctgaggcg gaaagaacca gtggcggtaa tacggttatc cacagaatca
     4201 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4261 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4321 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4381 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4441 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4501 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4561 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4621 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4681 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     4741 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4801 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4861 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4921 tcacgttaag ggattttggt catgacatta acctataaaa ataggcgtat cacgaggccc
     4981 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     5041 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     5101 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     5161 agagtgcacc atatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     5221 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     5281 cttcgctatt acgcca
//
