LOCUS       Exported                5851 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    HA-VHL Y112H-pBabe-puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5851)
  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 5851)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5851
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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(2090..2110)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2095..2424
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2275..2410
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2337..2356
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2434..3033
                     /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(2434..2453)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2930..2950
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3132..3725
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3926..3943)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4097..4685)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4177..4196)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4863..5723)
                     /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     5486..5505
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5724..5828)
                     /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 cggatccacc atgccccgga gggcggagaa
     1441 ctgggacgag gccgaggtag gcgcggagga ggcaggcgtc gaagagtacg gccctgaaga
     1501 agacggcggg gaggagtcgg gcgccgagga gtccggcccg gaagagtccg gcccggagga
     1561 actgggcgcc gaggaggaga tggaggccgg gcggccgcgg cccgtgctgc gctcggtgaa
     1621 ctcgcgcgag ccctcccagg tcatcttctg caatcgcagt ccgcgcgtcg tgctgcccgt
     1681 atggctcaac ttcgacggcg agccgcagcc ctacccaacg ctgccgcctg gcacgggccg
     1741 ccgcatccac agccaccgag gtcacctttg gctcttcaga gatgcaggga cacacgatgg
     1801 gcttctggtt aaccaaactg aattatttgt gccatctctc aatgttgacg gacagcctat
     1861 ttttgccaat atcacactgc cagtgtatac tctgaaagag cgatgcctcc aggttgtccg
     1921 gagcctagtc aagcctgaga attacaggag actggacatc gtcaggtcgc tctacgaaga
     1981 tctggaagac cacccaaatg tgcagaaaga cctggagcgg ctgacacagg agcgcattgc
     2041 acatcaacgg atgggagatt gaattcgcca gcacagtggt cgaccctgtg gaatgtgtgt
     2101 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2161 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2221 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     2281 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     2341 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     2401 tttggaggcc taggcttttg caaaaagctt accatgaccg agtacaagcc cacggtgcgc
     2461 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     2521 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     2581 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     2641 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     2701 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     2761 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     2821 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     2881 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     2941 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg agtgcccgaa ggaccgcgcg
     3001 acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg
     3061 accgaaagga gcgcacgacc ccatgcatcg ataaaataaa agattttatt tagtctccag
     3121 aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca
     3181 ttttgcaagg catggaaaaa tacataactg agaatagaga agttcagatc aaggtcagga
     3241 acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
     3301 ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat atctgtggta
     3361 agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg gtccagccct
     3421 cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc
     3481 tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct
     3541 ccccgagctc aataaaagag cccacaaccc ctcactcggg gcgccagtcc tccgattgac
     3601 tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc cgacttgtgg
     3661 tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt
     3721 tcatttccga cttgtggtct cgctgccttg ggagggtctc ctctgagtga ttgactaccc
     3781 gtcagcgggg gtcttcacat gcagcatgta tcaaaattaa tttggttttt tttcttaagt
     3841 atttacatta aatggccata gttgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     3901 ttgcgtattg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3961 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4021 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4081 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4141 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4201 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4261 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4321 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4381 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4441 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4501 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     4561 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4621 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4681 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4741 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     4801 aaaaatgaag tttgcggccg caaatcaatc taaagtatat atgagtaaac ttggtctgac
     4861 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4921 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4981 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5041 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5101 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5161 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5221 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5281 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5341 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5401 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5461 tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg
     5521 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5581 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5641 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5701 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5761 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5821 gttccgcgca catttccccg aaaagtgcca c
//
