LOCUS       Exported                6308 bp ds-DNA     circular SYN 04-JUL-2014
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pD164-pBABE.puro_TO_Flag_HuR.wt
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6308)
  AUTHORS   Douglas Adamoski
  TITLE     Direct Submission
  JOURNAL   Exported quarta-feira, 4 de abr de 2018 from SnapGene 2.8.3
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6308
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     LTR             8..479
                     /note="LTR (truncated)"
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    541
                     /note="splice donor mutation"
                     /note="T-to-C mutation that blocks RNA splicing"
     misc_feature    542..741
                     /note="MMLV Psi"
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             942..1358
                     /note="gag (truncated)"
                     /note="truncated MMLV gag gene lacking the start codon"
     misc_feature    1363
                     /note="MCS"
                     /note="multiple cloning site"
     promoter        1364..1420
                     /note="CMV Promoter"
     protein_bind    1364..1365
                     /note="TetO2"
     protein_bind    1368..1386
                     /note="TetO2"
     -10_signal      1513..1518
                     /note="Kozak"
     CDS             1516..1542
                     /codon_start=1
                     /note="Flag"
                     /translation="MDYKDDDDK"
     CDS             1555..2535
                     /codon_start=1
                     /gene="ELAVL"
                     /note="HuR (wt)"
                     /note="NM_001419.2"
                     /translation="MSNGYEDHMAEDCRGDIGRTNLIVNYLPQNMTQDELRSLFSSIGE
                     VESAKLIRDKVAGHSLGYGFVNYVTAKDAERAINTLNGLRLQSKTIKVSYARPSSEVIK
                     DANLYISGLPRTMTQKDVEDMFSRFGRIINSRVLVDQTTGLSRGVAFIRFDKRSEAEEA
                     ITSFNGHKPPGSSEPITVKFAANPNQNKNVALLSQLYHSPARRFGGPVHHQAQRFRFSP
                     MGVDHMSGLSGVNVPGNASSGWCIFIYNLGQDADEGILWQMFGPFGAVTNVKVIRDFNT
                     NKCKGFGFVTMTNYEEAAMAIASLNGYRLGDKILQVSFKTNKSHK"
     misc_feature    2537..2541
                     /note="MCS"
                     /note="multiple cloning site"
     promoter        2552..2881
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      2732..2867
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     CDS             2891..3490
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /note="PuroR"
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     LTR             3589..4182
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     rep_origin      complement(4554..5142)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5320..6180)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6181..6285)
                     /gene="bla"
                     /note="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 caagctggcc 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 tgctgcagcg ctgcagcatc gttctgtgtt
      901 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
      961 ccttaagttt gaccttagat cactggaaag atgtcgagcg gctcgctcac aaccagtcgg
     1021 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     1081 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     1141 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     1201 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     1261 ctcttcttcc atccgcgccg tctctccccc ttgaacctcc tctttcgacc ccgcctcaat
     1321 cctcccttta tccagccctc actccttctc taggcgccgg ccggatctcc ctatcagtga
     1381 tagagatcgt cgacgagctc gtttagtgaa ccgtcagatc gcctggagac gccatccacg
     1441 ctgttttgac ctccatagaa gacaccggga ccgatccagc ctccggactc tagcgtttaa
     1501 acttaagctg ccaccatgga ctacaaggac gacgatgaca agaagcttgg taccatgtct
     1561 aatggttatg aagaccacat ggccgaagac tgcaggggtg acatcgggag aacgaatttg
     1621 atcgtcaact acctccctca gaacatgacc caggatgagt tacgaagcct gttcagcagc
     1681 attggtgaag ttgaatctgc aaaacttatt cgggataaag tagcaggaca cagcttgggc
     1741 tatggctttg tgaactacgt gaccgcgaag gatgcagaga gagcgatcaa cacgctgaac
     1801 ggcttgaggc tccagtcaaa aaccattaag gtgtcgtatg ctcgcccgag ctcagaggtg
     1861 atcaaagacg ccaacttgta catcagcggg ctcccgcgga ccatgaccca gaaggacgta
     1921 gaagacatgt tctctcggtt tgggcggatc atcaactcgc gggtcctcgt ggatcagact
     1981 acaggtttgt ccagaggggt tgcgtttatc cggtttgaca aacggtcgga ggcagaagag
     2041 gcaattacca gtttcaatgg tcataaaccc ccaggttcct ctgagcccat cacagtgaag
     2101 tttgcagcca accccaacca gaacaaaaac gtggcactcc tctcgcagct gtaccactcg
     2161 ccagcgcgac ggttcggagg ccccgttcac caccaggcgc agagattcag gttctccccc
     2221 atgggcgtcg atcacatgag cgggctctct ggcgtcaacg tgccaggaaa cgcctcctcc
     2281 ggctggtgca ttttcatcta caacctgggg caggatgccg acgaggggat cctctggcag
     2341 atgtttgggc cgtttggtgc cgtcaccaat gtgaaagtga tccgcgactt caacaccaac
     2401 aagtgcaaag ggtttggctt tgtgaccatg acaaactatg aagaagccgc gatggccata
     2461 gccagcctga acggctaccg cctgggggac aaaatcttac aggtttcctt caaaaccaac
     2521 aagtcccaca aataactcga ccctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     2581 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     2641 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     2701 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     2761 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     2821 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     2881 aaagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     2941 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     3001 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     3061 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     3121 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     3181 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     3241 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
     3301 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     3361 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     3421 cgtcaccgcc gacgtcgagt gcccgaagga ccgcgcgacc tggtgcatga cccgcaagcc
     3481 cggtgcctga cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg cacgacccca
     3541 tgcatcgata aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc
     3601 cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaaatac
     3661 ataactgaga atagagaagt tcagatcaag gtcaggaaca gatggaacag ctgaatatgg
     3721 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
     3781 aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg
     3841 gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag agaaccatca
     3901 gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa
     3961 tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc
     4021 acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg tacccgtgta
     4081 tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc
     4141 tcctctgagt gattgactac ccgtcagcgg gggtctttca tttccgactt gtggtctcgc
     4201 tgccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc ttcacatgca
     4261 gcatgtatca aaattaattt ggtttttttt cttaagtatt tacattaaat ggccatagtt
     4321 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattggcg ctcttccgct
     4381 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4441 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4501 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4561 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4621 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4681 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4741 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4801 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4861 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4921 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4981 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5041 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5101 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5161 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5221 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt gcggccgcaa
     5281 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5341 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5401 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5461 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5521 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5581 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5641 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5701 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5761 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5821 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5881 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg
     5941 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6001 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6061 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6121 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6181 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6241 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6301 agtgccac
//
