LOCUS       sequence-12249-e        7844 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 7844)
  AUTHORS   Kondo K, Klco J, Nakamura E, Lechpammer M, Kaelin WG
  TITLE     Inhibition of HIF is necessary for tumor suppression by the von 
            Hippel-Lindau protein.
  JOURNAL   Cancer Cell. 2002 Apr . 1(3):237-46.
  PUBMED    12086860
REFERENCE   2  (bases 1 to 7844)
  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 
            Cell. 2002 Apr . 1(3):237-46."
COMMENT     Sequence Label: HA-HIF2-alpha P531A-pBabe-puro
FEATURES             Location/Qualifiers
     source          1..7844
                     /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(4083..4103)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4088..4417
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4268..4403
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4427..5026
                     /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(4427..4446)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4923..4943
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5125..5718
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5919..5936)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6090..6678)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6170..6189)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6856..7647,7648..7716))
                     /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     7479..7498
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7717..7821)
                     /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 accatgacag ctgacaagga
     1441 gaagaaaagg agtagctcgg agaggaggaa ggagaagtcc cgggatgctg cgcggtgccg
     1501 gcggagcaag gagacggagg tgttctatga gctggcccat gagctgcctc tgccccacag
     1561 tgtgagctcc catctggaca aggcctccat catgcgactg gcaatcagct tcctgcgaac
     1621 acacaagctc ctctcctcag tttgctctga aaacgagtcc gaagccgaag ctgaccagca
     1681 gatggacaac ttgtacctga aagccttgga gggtttcatt gccgtggtga cccaagatgg
     1741 cgacatgatc tttctgtcag aaaacatcag caagttcatg ggacttacac aggtggagct
     1801 aacaggacat agtatctttg acttcactca tccctgcgac catgaggaga ttcgtgagaa
     1861 cctgagtctc aaaaatggct ctggttttgg gaaaaaaagc aaagacatgt ccacagagcg
     1921 ggacttcttc atgaggatga agtgcacggt caccaacaga ggccgtactg tcaacctcaa
     1981 gtcagccacc tggaaggtct tgcactgcac gggccaggtg aaagtctaca acaactgccc
     2041 tcctcacaat agtctgtgtg gctacaagga gcccctgctg tcctgcctca tcatcatgtg
     2101 tgaaccaatc cagcacccat cccacatgga catccccctg gatagcaaga ccttcctgag
     2161 ccgccacagc atggacatga agttcaccta ctgtgatgac agaatcacag aactgattgg
     2221 ttaccaccct gaggagctgc ttggccgctc agcctatgaa ttctaccatg cgctagactc
     2281 cgagaacatg accaagagtc accagaactt gtgcaccaag ggtcaggtag taagtggcca
     2341 gtaccggatg ctcgcaaagc atgggggcta cgtgtggctg gagacccagg ggacggtcat
     2401 ctacaaccct cgcaacctgc agccccagtg catcatgtgt gtcaactacg tcctgagtga
     2461 gattgagaag aatgacgtgg tgttctccat ggaccagact gaatccctgt tcaagcccca
     2521 cctgatggcc atgaacagca tctttgatag cagtggcaag ggggctgtgt ctgagaagag
     2581 taacttccta ttcaccaagc taaaggagga gcccgaggag ctggcccagc tggctcccac
     2641 cccaggagac gccatcatct ctctggattt cgggaatcag aacttcgagg agtcctcagc
     2701 ctatggcaag gccatcctgc ccccgagcca gccatgggcc acggagttga ggagccacag
     2761 cacccagagc gaggctggga gcctgcctgc cttcaccgtg ccccaggcag ctgccccggg
     2821 cagcaccacc cccagtgcca ccagcagcag cagcagctgc tccacgccca atagccctga
     2881 agactattac acatctttgg ataacgacct gaagattgaa gtgattgaga agctcttcgc
     2941 catggacaca gaggccaagg accaatgcag tacccagacg gatttcaatg agctggactt
     3001 ggagacactg gcagctctat atccccatgg acggggaaga cttccagcta agccccatct
     3061 gccccgagga gcggctcttg gcggagaacc cacagtccac cccccagcac tgcttcagtg
     3121 ccatgacaaa catcttccag ccactggccc ctgtagcccc gcacagtccc ttcctcctgg
     3181 acaagtttca gcagcagctg gagagcaaga agacagagcc cgagcaccgg cccatgtcct
     3241 ccatcttctt tgatgccgga agcaaagcat ccctgccacc gtgctgtggc caggccagca
     3301 cccctctctc ttccatgggg ggcagatcca atacccagtg gcccccagat ccaccattac
     3361 attttgggcc cacaaagtgg gccgtcgggg atcagcgcac agagttcttg ggagcagcgc
     3421 cgttggggcc ccctgtctct ccaccccatg tctccacctt caagacaagg tctgcaaagg
     3481 gttttggggc tcgaggccca gacgtgctga gtccggccat ggtagccctc tccaacaagc
     3541 tgaagctgaa gcgacagctg gagtatgaag agcaagcctt ccaggacctg agcggggggg
     3601 acccacctgg tggcagcacc tcacatttga tgtggaaacg gatgaagaac ctcaggggtg
     3661 ggagctgccc tttgatgccg gacaagccac tgagcgcaaa tgtacccaat gataagttca
     3721 cccaaaaccc catgaggggc ctgggccatc ccctgagaca tctgccgctg ccacagcctc
     3781 catctgccat cagtcccggg gagaacagca agagcaggtt ccccccacag tgctacgcca
     3841 cccagtacca ggactacagc ctgtcgtcag cccacaaggt gtcaggcatg gcaagccggc
     3901 tgctcgggcc ctcatttgag tcctacctgc tgcccgaact gaccagatat gactgtgagg
     3961 tgaacgtgcc cgtgctggga agctccacgc tcctgcaagg aggggacctc ctcagagccc
     4021 tggaccaggc cacctgacaa ttcgatatct cgagtaatcg ccagcacagt ggtcgaccct
     4081 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4141 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4201 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4261 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4321 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     4381 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag cttaccatga ccgagtacaa
     4441 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc
     4501 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
     4561 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     4621 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     4681 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     4741 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     4801 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     4861 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     4921 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgagtgccc
     4981 gaaggaccgc gcgacctggt gcatgacccg caagcccggt gcctgacgcc cgccccacga
     5041 cccgcagcgc ccgaccgaaa ggagcgcacg accccatgca tcgataaaat aaaagatttt
     5101 atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc
     5161 ttaagtaacg ccattttgca aggcatggaa aaatacataa ctgagaatag agaagttcag
     5221 atcaaggtca ggaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag
     5281 cagttcctgc cccggctcag ggccaagaac agatggaaca gctgaatatg ggccaaacag
     5341 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat
     5401 gcggtccagc cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga
     5461 cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc
     5521 gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc ggggcgccag
     5581 tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc
     5641 atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt
     5701 cagcgggggt ctttcatttc cgacttgtgg tctcgctgcc ttgggagggt ctcctctgag
     5761 tgattgacta cccgtcagcg ggggtcttca catgcagcat gtatcaaaat taatttggtt
     5821 ttttttctta agtatttaca ttaaatggcc atagttgcat taatgaatcg gccaacgcgc
     5881 ggggagaggc ggtttgcgta ttggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5941 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     6001 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6061 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6121 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6181 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6241 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6301 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6361 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6421 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6481 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     6541 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6601 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6661 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6721 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6781 atccttttaa attaaaaatg aagtttgcgg ccgcaaatca atctaaagta tatatgagta
     6841 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6901 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6961 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7021 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7081 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7141 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7201 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7261 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7321 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7381 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7441 gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag
     7501 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7561 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7621 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7681 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7741 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7801 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccac
//
