LOCUS       sequence-12177-e        7821 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7821)
  AUTHORS   Yan Q, Bartz S, Mao M, Li L, Kaelin WG
  TITLE     The hypoxia-inducible factor 2alpha N-terminal and C-terminal 
            transactivation domains cooperate to promote renal tumorigenesis in 
            vivo.
  JOURNAL   Mol Cell Biol. 2007 Mar . 27(6):2092-102.
  PUBMED    17220275
REFERENCE   2  (bases 1 to 7821)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell 
            Biol. 2007 Mar . 27(6):2092-102."
COMMENT     Sequence Label: HA-HIF2-alpha P405A/P531A/N847A-pBabe-Puro
FEATURES             Location/Qualifiers
     source          1..7821
                     /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
                     /translation="YPYDVPDYA"
     primer_bind     complement(4060..4080)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4065..4394
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4245..4380
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4404..5003
                     /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(4404..4423)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4900..4920
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5102..5695
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5896..5913)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6067..6655)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6147..6166)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6833..7624,7625..7693))
                     /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     7456..7475
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7694..7798)
                     /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 cccctatgat gttcccgatt atgctggatc cgccaccatg acagctgaca aggagaagaa
     1441 aaggagtagc tcggagagga ggaaggagaa gtcccgggat gctgcgcggt gccggcggag
     1501 caaggagacg gaggtgttct atgagctggc ccatgagctg cctctgcccc acagtgtgag
     1561 ctcccatctg gacaaggcct ccatcatgcg actggcaatc agcttcctgc gaacacacaa
     1621 gctcctctcc tcagtttgct ctgaaaacga gtccgaagcc gaagctgacc agcagatgga
     1681 caacttgtac ctgaaagcct tggagggttt cattgccgtg gtgacccaag atggcgacat
     1741 gatctttctg tcagaaaaca tcagcaagtt catgggactt acacaggtgg agctaacagg
     1801 acatagtatc tttgacttca ctcatccctg cgaccatgag gagattcgtg agaacctgag
     1861 tctcaaaaat ggctctggtt ttgggaaaaa aagcaaagac atgtccacag agcgggactt
     1921 cttcatgagg atgaagtgca cggtcaccaa cagaggccgt actgtcaacc tcaagtcagc
     1981 cacctggaag gtcttgcact gcacgggcca ggtgaaagtc tacaacaact gccctcctca
     2041 caatagtctg tgtggctaca aggagcccct gctgtcctgc ctcatcatca tgtgtgaacc
     2101 aatccagcac ccatcccaca tggacatccc cctggatagc aagaccttcc tgagccgcca
     2161 cagcatggac atgaagttca cctactgtga tgacagaatc acagaactga ttggttacca
     2221 ccctgaggag ctgcttggcc gctcagccta tgaattctac catgcgctag actccgagaa
     2281 catgaccaag agtcaccaga acttgtgcac caagggtcag gtagtaagtg gccagtaccg
     2341 gatgctcgca aagcatgggg gctacgtgtg gctggagacc caggggacgg tcatctacaa
     2401 ccctcgcaac ctgcagcccc agtgcatcat gtgtgtcaac tacgtcctga gtgagattga
     2461 gaagaatgac gtggtgttct ccatggacca gactgaatcc ctgttcaagc cccacctgat
     2521 ggccatgaac agcatctttg atagcagtgg caagggggct gtgtctgaga agagtaactt
     2581 cctattcacc aagctaaagg aggagcccga ggagctggcc cagctggctg ctaccccagg
     2641 agacgccatc atctctctgg atttcgggaa tcagaacttc gaggagtcct cagcctatgg
     2701 caaggccatc ctgcccccga gccagccatg ggccacggag ttgaggagcc acagcaccca
     2761 gagcgaggct gggagcctgc ctgccttcac cgtgccccag gcagctgccc cgggcagcac
     2821 cacccccagt gccaccagca gcagcagcag ctgctccacg cccaatagcc ctgaagacta
     2881 ttacacatct ttggataacg acctgaagat tgaagtgatt gagaagctct tcgccatgga
     2941 cacagaggcc aaggaccaat gcagtaccca gacggatttc aatgagctgg acttggagac
     3001 actggcagct tatatcccca tggacgggga agacttccag ctaagcccca tctgccccga
     3061 ggagcggctc ttggcggaga acccacagtc caccccccag cactgcttca gtgccatgac
     3121 aaacatcttc cagccactgg cccctgtagc cccgcacagt cccttcctcc tggacaagtt
     3181 tcagcagcag ctggagagca agaagacaga gcccgagcac cggcccatgt cctccatctt
     3241 ctttgatgcc ggaagcaaag catccctgcc accgtgctgt ggccaggcca gcacccctct
     3301 ctcttccatg gggggcagat ccaataccca gtggccccca gatccaccat tacattttgg
     3361 gcccacaaag tgggccgtcg gggatcagcg cacagagttc ttgggagcag cgccgttggg
     3421 gccccctgtc tctccacccc atgtctccac cttcaagaca aggtctgcaa agggttttgg
     3481 ggctcgaggc ccagacgtgc tgagtccggc catggtagcc ctctccaaca agctgaagct
     3541 gaagcgacag ctggagtatg aagagcaagc cttccaggac ctgagcgggg gggacccacc
     3601 tggtggcagc acctcacatt tgatgtggaa acggatgaag aacctcaggg gtgggagctg
     3661 ccctttgatg ccggacaagc cactgagcgc aaatgtaccc aatgataagt tcacccaaaa
     3721 ccccatgagg ggcctgggcc atcccctgag acatctgccg ctgccacagc ctccatctgc
     3781 catcagtccc ggggagaaca gcaagagcag gttcccccca cagtgctacg ccacccagta
     3841 ccaggactac agcctgtcgt cagcccacaa ggtgtcaggc atggcaagcc ggctgctcgg
     3901 gccctcattt gagtcctacc tgctgcccga actgaccaga tatgactgtg aggtgaacgt
     3961 gcccgtgctg ggaagctcca cgctcctgca aggaggggac ctcctcagag ccctggacca
     4021 ggccacctga caattcgcca gcacagtggt cgaccctgtg gaatgtgtgt cagttagggt
     4081 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4141 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4201 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4261 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4321 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4381 taggcttttg caaaaagctt accatgaccg agtacaagcc cacggtgcgc ctcgccaccc
     4441 gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca
     4501 cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct
     4561 tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg
     4621 tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc
     4681 cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc
     4741 tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg
     4801 accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc
     4861 gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc
     4921 ggctcggctt caccgtcacc gccgacgtcg agtgcccgaa ggaccgcgcg acctggtgca
     4981 tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg accgaaagga
     5041 gcgcacgacc ccatgcatcg ataaaataaa agattttatt tagtctccag aaaaaggggg
     5101 gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg
     5161 catggaaaaa tacataactg agaatagaga agttcagatc aaggtcagga acagatggaa
     5221 cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc
     5281 caagaacaga tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct
     5341 gccccggctc agggccaaga acagatggtc cccagatgcg gtccagccct cagcagtttc
     5401 tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat
     5461 ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc
     5521 aataaaagag cccacaaccc ctcactcggg gcgccagtcc tccgattgac tgagtcgccc
     5581 gggtacccgt gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt
     5641 ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt tcatttccga
     5701 cttgtggtct cgctgccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg
     5761 gtcttcacat gcagcatgta tcaaaattaa tttggttttt tttcttaagt atttacatta
     5821 aatggccata gttgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5881 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5941 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6001 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6061 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6121 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6181 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6241 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6301 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6361 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6421 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6481 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     6541 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6601 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6661 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6721 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6781 tttgcggccg caaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6841 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6901 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6961 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7021 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7081 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7141 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7201 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7261 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7321 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7381 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7441 cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7501 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7561 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7621 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7681 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7741 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7801 catttccccg aaaagtgcca c
//
