LOCUS       sequence-242748-        5267 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 5267)
  AUTHORS   Shiao YH, Resau JH, Nagashima K, Anderson LM, Ramakrishna G
  TITLE     The von Hippel-Lindau tumor suppressor targets to mitochondria.
  JOURNAL   Cancer Res. 2000 Jun 1. 60(11):2816-9.
  PUBMED    10850420
REFERENCE   2  (bases 1 to 5267)
  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: "Cancer Res.
            2000 Jun 1. 60(11):2816-9."
COMMENT     Sequence Label: pEGFP-N1-wtVHL
FEATURES             Location/Qualifiers
     source          1..5267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        85..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        389..592
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     543..563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1231..1240
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1237..1239,1240..1242,1243..1956)
                     /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(1237..1239,1240..1242,1243..1953)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1282..1303)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1543..1562)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1890..1911
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2079..2200
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2116..2135)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2170..2189
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2207..2662)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2344..2365)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2556..2575
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2689..2793
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2795..3152
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2795..2986
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2818..2838)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3003..3138
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3187..3981
                     /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             3187..3981
                     /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(3241..3260)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3851..3870
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4169..4188)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4213..4260
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4589..5177
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5078..5097
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt
       61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      361 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      421 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      481 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      541 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
      601 gaaccgtcag atccgctagc gctaccggac tcagatctcg agctcaagct tcggccacca
      661 tgccccggaa ggcagctagt ccagaggagg cagaaaggat gccgggctct gaagagatag
      721 aggctgggcg gccgcggccg gttttacgct ctgtgaactc gcgcgaaccc tctcaggtca
      781 tcttctgcaa ccgcagcccg cgcgtcgtgc tgcctttgtg gctcaacttt gatggtgagc
      841 ctcagcccta cccgacctta ccaccgggca ccggccgccg catccacagc taccgaggtc
      901 acctttggct cttcagggat gcggggaccc atgatggact tctggttaac caaacggaac
      961 tgtttgtgcc atccctcaat gttgatggac agcctatttt tgccaacatc acattgccag
     1021 tgtataccct gaaagagcgg tgccttcagg ttgtacggag cctggtcaag cctgagaact
     1081 acaggaggct ggacatcgtc aggtcgctct atgaagactt ggaagaccac ccaaatgtgc
     1141 ggaaagacat acagcggctg acccaagagc acctcgagaa tcaggccctg ggagaggagc
     1201 ctgaaggagt ccactgagga tccaccggtc gccaccatgg tgagcaaggg cgaggagctg
     1261 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     1321 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     1381 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     1441 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     1501 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     1561 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     1621 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     1681 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     1741 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     1801 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     1861 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     1921 gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatcataa
     1981 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     2041 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     2101 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     2161 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt
     2221 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     2281 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     2341 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     2401 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     2461 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     2521 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     2581 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     2641 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2701 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2761 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     2821 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2881 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     2941 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3001 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3061 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3121 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     3181 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3241 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3301 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3361 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3421 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3481 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3541 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3601 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3661 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     3721 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     3781 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     3841 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     3901 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     3961 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4021 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4081 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4141 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     4201 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     4261 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     4321 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     4381 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     4441 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4501 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4561 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4621 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4681 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4741 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4801 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4861 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4921 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4981 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5041 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5101 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5161 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5221 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtg
//
