LOCUS       sequence-11270-e        5053 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 5053)
  AUTHORS   Beskow A, Grimberg KB, Bott LC, Salomons FA, Dantuma NP, Young P
  TITLE     A conserved unfoldase activity for the p97 AAA-ATPase in proteasomal
            degradation.
  JOURNAL   J Mol Biol. 2009 Dec 11. 394(4):732-46.
  PUBMED    19782090
REFERENCE   2  (bases 1 to 5053)
  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: "J Mol Biol.
            2009 Dec 11. 394(4):732-46."
COMMENT     Sequence Label: UbG76V-eGFP-V5His_20aa
FEATURES             Location/Qualifiers
     source          1..5053
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    120..241
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(157..176)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     211..230
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(248..703)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(385..406)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     597..616
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        730..834
                     /gene="bla"
                     /label=AmpR promoter
     promoter        836..1193
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        836..1027
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(859..879)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1044..1179
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1228..2022
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1228..2022
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1282..1301)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1892..1911
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2210..2229)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2254..2301
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2630..3218
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3119..3138
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3393..3696
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3697..3900
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3851..3871
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3960..4184
                     /codon_start=1
                     /product="human ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRG"
     regulatory      4221..4230
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4227..4229,4230..4232,4233..4943)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4227..4229,4230..4232,4233..4943)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4272..4293)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4533..4552)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4880..4901
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5031..5048
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa
       61 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
      121 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
      181 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
      241 aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
      301 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
      361 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
      421 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
      481 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
      541 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
      601 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
      661 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg
      721 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      781 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag
      841 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
      901 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
      961 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     1021 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     1081 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     1141 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat
     1201 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     1261 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     1321 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     1381 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc
     1441 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     1501 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     1561 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     1621 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     1681 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     1741 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     1801 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     1861 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     1921 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     1981 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     2041 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     2101 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     2161 gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga
     2221 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     2281 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     2341 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     2401 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     2461 ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat ttttaattta
     2521 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     2581 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     2641 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     2701 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2761 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2821 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2881 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2941 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3001 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3061 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3121 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3181 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3241 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     3301 attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg
     3361 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     3421 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     3481 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     3541 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     3601 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     3661 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     3721 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     3781 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     3841 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     3901 ggtttagtga accgtcagat ccgctagcgc taccggactc agatctcgag ctcggatcca
     3961 tgcagatctt cgtgaagact ctgactggta agaccatcac cctcgaggtt gagcccagtg
     4021 acaccattga gaatgtcaag gcaaagatcc aagataagga aggcatccct cctgaccagc
     4081 agaggctgat ctttgctgga aaacagctgg aagatgggcg caccctgtct gactacaaca
     4141 tccagaaaga gtccaccctg cacctggtac tccgtctcag aggtgtggtg gggaagcttg
     4201 gtcgacagga tccaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     4261 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     4321 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     4381 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     4441 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     4501 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     4561 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     4621 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     4681 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     4741 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     4801 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     4861 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     4921 tcggcatgga cgagctgtac aagtcatgtc ggccgctcga gtctagaggg cccttcgaag
     4981 gtaagcctat ccctaaccct ctctgaggtc tcgattctac gcgtaccggt catcatcacc
     5041 atcaccattg agc
//
