LOCUS       Exported                5791 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    HA-VHL wt-pBabe-puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5791)
  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 5791)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5791
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             6..475
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    540..739
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             940..1356
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1286..1308
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1324..1340
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1388..1414
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2119..2139)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2124..2453
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2304..2439
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2366..2385
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2463..3062
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2463..2482)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2959..2979
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3283..3752
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3882..3899)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4053..4641)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4133..4152)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4801..5661)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5424..5443
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5662..5766)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gcagcctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg
       61 gccaagaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc
      121 ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtccagcc ctcagcagtt
      181 tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt
      241 atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc
      301 tcaataaaag agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc
      361 ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg
      421 ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatttgg
      481 gggctcgtcc gggatcggga gacccctgcc cagggaccac cgacccacca ccgggaggca
      541 agctggccag caacttatct gtgtctgtcc gattgtctag tgtctatgac tgattttatg
      601 cgcctgcgtc ggtactagtt agctaactag ctctgtatct ggcggacccg tggtggaact
      661 gacgagttct gaacacccgg ccgcaaccct gggagacgtc ccagggactt tgggggccgt
      721 ttttgtggcc cgacctgagg aagggagtcg atgtggaatc cgaccccgtc aggatatgtg
      781 gttctggtag gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg
      841 gtttggaacc gaagccgcgc gtcttgtctg ctgcagcgct gcagcatcgt tctgtgttgt
      901 ctctgtctga ctgtgtttct gtatttgtct gaaaattagg gccagactgt taccactccc
      961 ttaagtttga ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa ccagtcggta
     1021 gatgtcaaga agagacgttg ggttaccttc tgctctgcag aatggccaac ctttaacgtc
     1081 ggatggccgc gagacggcac ctttaaccga gacctcatca cccaggttaa gatcaaggtc
     1141 ttttcacctg gcccgcatgg acacccagac caggtcccct acatcgtgac ctgggaagcc
     1201 ttggcttttg acccccctcc ctgggtcaag ccctttgtac accctaagcc tccgcctcct
     1261 cttcctccat ccgccccgtc tctccccctt gaacctcctc gttcgacccc gcctcgatcc
     1321 tccctttatc cagccctcac tccttctcta ggcgccggcc ggatcgaaga ctaagcttac
     1381 catggcctac ccctacgacg tgcccgacta cgcctccctc ggatccgcca ccatgccccg
     1441 gagggcggag aactgggacg aggccgaggt aggcgcggag gaggcaggcg tcgaagagta
     1501 cggccctgaa gaagacggcg gggaggagtc gggcgccgag gagtccggcc cggaagagtc
     1561 cggcccggag gaactgggcg ccgaggagga gatggaggcc gggcggccgc ggcccgtgct
     1621 gcgctcggtg aactcgcgcg agccctccca ggtcatcttc tgcaatcgca gtccgcgcgt
     1681 cgtgctgccc gtatggctca acttcgacgg cgagccgcag ccctacccaa cgctgccgcc
     1741 tggcacgggc cgccgcatcc acagctaccg aggtcacctt tggctcttca gagatgcagg
     1801 gacacacgat gggcttctgg ttaaccaaac tgaattattt gtgccatctc tcaatgttga
     1861 cggacagcct atttttgcca atatcacact gccagtgtat actctgaaag agcgatgcct
     1921 ccaggttgtc cggagcctag tcaagcctga gaattacagg agactggaca tcgtcaggtc
     1981 gctctacgaa gatctggaag accacccaaa tgtgcagaaa gacctggagc ggctgacaca
     2041 ggagcgcatt gcacatcaac ggatgggaga ttgaattcga tatctcgagt aattcgccag
     2101 cacagtggtc gaccctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     2161 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     2221 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     2281 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     2341 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     2401 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctta
     2461 ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg
     2521 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg
     2581 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     2641 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     2701 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     2761 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     2821 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     2881 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     2941 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     3001 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     3061 gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catgcatcga
     3121 taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta
     3181 ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaaat acataactga
     3241 gaatagagaa gttcagatca aggtcaggaa cagatggaac agctgaatat gggccaaaca
     3301 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggaacagctg
     3361 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     3421 cagatggtcc ccagatgcgg tccagccctc agcagtttct agagaaccat cagatgtttc
     3481 cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg
     3541 cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc
     3601 tcactcgggg cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa
     3661 accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga
     3721 gtgattgact acccgtcagc gggggtcttt cacatgcagc atgtatcaaa attaatttgg
     3781 ttttttttct taagtattta cattaaatgg ccatagttgc attaatgaat cggccaacgc
     3841 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     3901 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3961 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4021 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4081 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4141 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4201 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4261 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4321 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4381 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4441 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4501 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4561 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4621 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4681 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4741 tagatccttt tgcggccgca aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4801 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4861 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4921 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4981 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5041 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5101 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5161 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5221 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5281 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5341 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5401 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5461 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5521 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5581 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5641 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5701 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5761 tccgcgcaca tttccccgaa aagtgccacc t
//
