LOCUS       sequence-8957-ex        5854 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 5854)
  AUTHORS   Li L, Zhang L, Zhang X, Yan Q, Minamishima YA, Olumi AF, Mao M, 
            Bartz S, Kaelin WG
  TITLE     Hypoxia-inducible factor linked to differential kidney cancer risk 
            seen with type 2A and type 2B VHL mutations.
  JOURNAL   Mol Cell Biol. 2007 Aug . 27(15):5381-92.
  PUBMED    17526729
REFERENCE   2  (bases 1 to 5854)
  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: "Mol Cell 
            Biol. 2007 Aug . 27(15):5381-92."
COMMENT     Sequence Label: HA-VHL W117R-pBabe-puro
FEATURES             Location/Qualifiers
     source          1..5854
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             8..477
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    542..741
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             934..1350
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1318..1334
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1379..1405
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             1379..1405
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1385..1411
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     primer_bind     complement(2093..2113)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2098..2427
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2278..2413
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2437..3036
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     primer_bind     complement(2437..2456)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2933..2953
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3135..3728
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3929..3946)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4100..4688)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4180..4199)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4866..5657,5658..5726))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5489..5508
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5727..5831)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
       61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
      121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
      181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
      241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
      301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
      361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
      421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
      481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
      541 taagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
      601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
      661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
      721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
      781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
      841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt
      901 ctgactgtgt ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt
      961 ttgaccttag atcactggaa agatgtcgag cggctcgctc acaaccagtc ggtagatgtc
     1021 aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg
     1081 ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca
     1141 cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct
     1201 tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttctt
     1261 ccatccgcgc cgtctctccc ccttgaacct cctctttcga ccccgcctca atcctccctt
     1321 tatccagccc tcactccttc tctaggcgcc ggccatcgag actaagctta ccatggccta
     1381 cccctacgac gtgcccgact acgcctccct cggatccgcc accatgcccc ggagggcgga
     1441 gaactgggac gaggccgagg taggcgcgga ggaggcaggc gtcgaagagt acggccctga
     1501 agaagacggc ggggaggagt cgggcgccga ggagtccggc ccggaagagt ccggcccgga
     1561 ggaactgggc gccgaggagg agatggaggc cgggcggccg cggcccgtgc tgcgctcggt
     1621 gaactcgcgc gagccctccc aggtcatctt ctgcaatcgc agtccgcgcg tcgtgctgcc
     1681 cgtatggctc aacttcgacg gcgagccgca gccctaccca acgctgccgc ctggcacggg
     1741 ccgccgcatc cacagctacc gaggtcacct tcggctcttc agagatgcag ggacacacga
     1801 tgggcttctg gttaaccaaa ctgaattatt tgtgccatct ctcaatgttg acggacagcc
     1861 tatttttgcc aatatcacac tgccagtgta tactctgaaa gagcgatgcc tccaggttgt
     1921 ccggagccta gtcaagcctg agaattacag gagactggac atcgtcaggt cgctctacga
     1981 agatctggaa gaccacccaa atgtgcagaa agacctggag cggctgacac aggagcgcat
     2041 tgcacatcaa cggatgggag attgaattcg ccagcacagt ggtcgaccct gtggaatgtg
     2101 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2161 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     2221 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     2281 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     2341 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     2401 ttttttggag gcctaggctt ttgcaaaaag cttaccatga ccgagtacaa gcccacggtg
     2461 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc
     2521 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag
     2581 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac
     2641 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc
     2701 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag
     2761 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc
     2821 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg
     2881 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc
     2941 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgagtgccc gaaggaccgc
     3001 gcgacctggt gcatgacccg caagcccggt gcctgacgcc cgccccacga cccgcagcgc
     3061 ccgaccgaaa ggagcgcacg accccatgca tcgataaaat aaaagatttt atttagtctc
     3121 cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg
     3181 ccattttgca aggcatggaa aaatacataa ctgagaatag agaagttcag atcaaggtca
     3241 ggaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc
     3301 cccggctcag ggccaagaac agatggaaca gctgaatatg ggccaaacag gatatctgtg
     3361 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtccagc
     3421 cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga
     3481 ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct
     3541 gctccccgag ctcaataaaa gagcccacaa cccctcactc ggggcgccag tcctccgatt
     3601 gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg
     3661 tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt
     3721 ctttcatttc cgacttgtgg tctcgctgcc ttgggagggt ctcctctgag tgattgacta
     3781 cccgtcagcg ggggtcttca catgcagcat gtatcaaaat taatttggtt ttttttctta
     3841 agtatttaca ttaaatggcc atagttgcat taatgaatcg gccaacgcgc ggggagaggc
     3901 ggtttgcgta ttggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3961 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4021 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4081 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4141 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4201 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4261 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4321 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4381 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4441 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4501 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     4561 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4621 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4681 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4741 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4801 attaaaaatg aagtttgcgg ccgcaaatca atctaaagta tatatgagta aacttggtct
     4861 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4921 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4981 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5041 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5101 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5161 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5221 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5281 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5341 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5401 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5461 agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa
     5521 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5581 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5641 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5701 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5761 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5821 ggggttccgc gcacatttcc ccgaaaagtg ccac
//
