LOCUS       sequence-10153-e        7843 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 7843)
  AUTHORS   Kondo K, Kim WY, Lechpammer M, Kaelin WG
  TITLE     Inhibition of HIF2alpha is sufficient to suppress pVHL-defective 
            tumor growth.
  JOURNAL   PLoS Biol. 2003 Dec . 1(3):E83.
  PUBMED    14691554
REFERENCE   2  (bases 1 to 7843)
  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 Biol. 
            2003 Dec . 1(3):E83."
COMMENT     Sequence Label: HA-HIF2-alpha P405A;P531A;bHLH*-pBabe-Puro
FEATURES             Location/Qualifiers
     source          1..7843
                     /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(4082..4102)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4087..4416
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4267..4402
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4426..5025
                     /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(4426..4445)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4922..4942
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5124..5717
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5918..5935)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6089..6677)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6169..6188)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6855..7646,7647..7715))
                     /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     7478..7497
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7716..7820)
                     /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 cggcttgcgc
     1501 tgctagcgct gagacggagg tgttctatga gctggcccat gagctgcctc tgccccacag
     1561 tgtgagctcc catctggaca aggcctccat catgcgactg gaaatcagct 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 gcaccctata tccccatgga cggggaagac ttccagctaa gccccatctg
     3061 ccccgaggag cggctcttgg cggagaaccc acagtccacc ccccagcact gcttcagtgc
     3121 catgacaaac atcttccagc cactggcccc tgtagccccg cacagtccct tcctcctgga
     3181 caagtttcag cagcagctgg agagcaagaa gacagagccc gagcaccggc ccatgtcctc
     3241 catcttcttt gatgccggaa gcaaagcatc cctgccaccg tgctgtggcc aggccagcac
     3301 ccctctctct tccatggggg gcagatccaa tacccagtgg cccccagatc caccattaca
     3361 ttttgggccc acaaagtggg ccgtcgggga tcagcgcaca gagttcttgg gagcagcgcc
     3421 gttggggccc cctgtctctc caccccatgt ctccaccttc aagacaaggt ctgcaaaggg
     3481 ttttggggct cgaggcccag acgtgctgag tccggccatg gtagccctct ccaacaagct
     3541 gaagctgaag cgacagctgg agtatgaaga gcaagccttc caggacctga gcggggggga
     3601 cccacctggt ggcagcacct cacatttgat gtggaaacgg atgaagaacc tcaggggtgg
     3661 gagctgccct ttgatgccgg acaagccact gagcgcaaat gtacccaatg ataagttcac
     3721 ccaaaacccc atgaggggcc tgggccatcc cctgagacat ctgccgctgc cacagcctcc
     3781 atctgccatc agtcccgggg agaacagcaa gagcaggttc cccccacagt gctacgccac
     3841 ccagtaccag gactacagcc tgtcgtcagc ccacaaggtg tcaggcatgg caagccggct
     3901 gctcgggccc tcatttgagt cctacctgct gcccgaactg accagatatg actgtgaggt
     3961 gaacgtgccc gtgctgggaa gctccacgct cctgcaagga ggggacctcc tcagagccct
     4021 ggaccaggcc acctgacaat tcgatatctc gagtaatcgc cagcacagtg gtcgaccctg
     4081 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4141 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4201 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4261 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4321 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     4381 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttaccatgac cgagtacaag
     4441 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     4501 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     4561 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     4621 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     4681 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     4741 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     4801 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     4861 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     4921 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgagtgcccg
     4981 aaggaccgcg cgacctggtg catgacccgc aagcccggtg cctgacgccc gccccacgac
     5041 ccgcagcgcc cgaccgaaag gagcgcacga ccccatgcat cgataaaata aaagatttta
     5101 tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct
     5161 taagtaacgc cattttgcaa ggcatggaaa aatacataac tgagaataga gaagttcaga
     5221 tcaaggtcag gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc
     5281 agttcctgcc ccggctcagg gccaagaaca gatggaacag ctgaatatgg gccaaacagg
     5341 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg
     5401 cggtccagcc ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac
     5461 ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg
     5521 cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg gggcgccagt
     5581 cctccgattg actgagtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca
     5641 tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc
     5701 agcgggggtc tttcatttcc gacttgtggt ctcgctgcct tgggagggtc tcctctgagt
     5761 gattgactac ccgtcagcgg gggtcttcac atgcagcatg tatcaaaatt aatttggttt
     5821 tttttcttaa gtatttacat taaatggcca tagttgcatt aatgaatcgg ccaacgcgcg
     5881 gggagaggcg gtttgcgtat tggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5941 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6001 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6061 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6121 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6181 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6241 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6301 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6361 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6421 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6481 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     6541 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6601 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6661 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6721 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6781 tccttttaaa ttaaaaatga agtttgcggc cgcaaatcaa tctaaagtat atatgagtaa
     6841 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6901 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6961 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     7021 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     7081 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     7141 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     7201 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     7261 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7321 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7381 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     7441 cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga
     7501 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7561 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7621 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7681 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7741 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7801 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cac
//
