LOCUS       Exported                7010 bp ds-DNA     circular SYN 04-ABR-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pD169-pBABE.puro_TO_Flag_tGFP.HuR.S221D
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7010)
  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..7010
                     /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"
     protein_bind    1364..1365
                     /note="TetO2"
     protein_bind    1368..1386
                     /note="TetO2"
     -10_signal      1513..1518
                     /note="Kozak"
     misc_signal     1516..1543
                     /note="MCS"
     CDS             1516..1542
                     /codon_start=1
                     /note="Flag"
                     /translation="MDYKDDDDK"
     CDS             1549..2244
                     /codon_start=1
                     /product="improved green fluorescent protein from 
                     Pontellina plumata (Evdokimov et al., 2006)"
                     /note="TurboGFP"
                     /note="mammalian codon-optimized"
                     /translation="MESDESGLPAMEIECRITGTLNGVEFELVGGGEGTPEQGRMTNKM
                     KSTKGALTFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVL
                     HVSFSYRYEAGRVIGDFKVMGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNDLDGSFTR
                     TFSLRDGGYYSSVVDSHMHFKSAIHPSILQNGGPMFAFRRVEEDHSNTELGIVEYQHAF
                     KTPDADAGEE"
     CDS             2257..3237
                     /codon_start=1
                     /gene="ELAVL"
                     /note="HuR (S221D)"
                     /note="NM_001419.2"
                     /translation="MSNGYEDHMAEDCRGDIGRTNLIVNYLPQNMTQDELRSLFSSIGE
                     VESAKLIRDKVAGHSLGYGFVNYVTAKDAERAINTLNGLRLQSKTIKVSYARPSSEVIK
                     DANLYISGLPRTMTQKDVEDMFSRFGRIINSRVLVDQTTGLSRGVAFIRFDKRSEAEEA
                     ITSFNGHKPPGSSEPITVKFAANPNQNKNVALLSQLYHSPARRFGGPVHHQAQRFRFDP
                     MGVDHMSGLSGVNVSGNASSGWCIFIYNLGQDADEGILWQMFGPFGAVTNVKVIRDFNT
                     NKCKGFGFVTMTNYEEAAMAIASLNGYRLGDKILQVSFKTNKSHK"
     variation       2917..2919
                     /note="S221D"
     variation       2965..2967
                     /note="P237S"
     promoter        3254..3583
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      3434..3569
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     CDS             3593..4192
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /note="PuroR"
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     LTR             4291..4884
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     rep_origin      complement(5256..5844)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6022..6882)
                     /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(6883..6987)
                     /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 agaagcttat ggagagcgac
     1561 gagagcggcc tgcccgccat ggagatcgag tgccgcatca ccggcaccct gaacggcgtg
     1621 gagttcgagc tggtgggcgg cggagagggc acccccgagc agggccgcat gaccaacaag
     1681 atgaagagca ccaaaggcgc cctgaccttc agcccctacc tgctgagcca cgtgatgggc
     1741 tacggcttct accacttcgg cacctacccc agcggctacg agaacccctt cctgcacgcc
     1801 atcaacaacg gcggctacac caacacccgc atcgagaagt acgaggacgg cggcgtgctg
     1861 cacgtgagct tcagctaccg ctacgaggcc ggccgcgtga tcggcgactt caaggtgatg
     1921 ggcaccggct tccccgagga cagcgtgatc ttcaccgaca agatcatccg cagcaacgcc
     1981 accgtggagc acctgcaccc catgggcgat aacgatctgg atggcagctt cacccgcacc
     2041 ttcagcctgc gcgacggcgg ctactacagc tccgtggtgg acagccacat gcacttcaag
     2101 agcgccatcc accccagcat cctgcagaac gggggcccca tgttcgcctt ccgccgcgtg
     2161 gaggaggatc acagcaacac cgagctgggc atcgtggagt accagcacgc cttcaagacc
     2221 ccggatgcag atgccggtga agaaagagtt ggtaccatgt ctaatggtta tgaagaccac
     2281 atggccgaag actgcagggg tgacatcggg agaacgaatt tgatcgtcaa ctacctccct
     2341 cagaacatga cccaggatga gttacgaagc ctgttcagca gcattggtga agttgaatct
     2401 gcaaaactta ttcgggataa agtagcagga cacagcttgg gctatggctt tgtgaactac
     2461 gtgaccgcga aggatgcaga gagagcgatc aacacgctga acggcttgag gctccagtca
     2521 aaaaccatta aggtgtcgta tgctcgcccg agctcagagg tgatcaaaga cgccaacttg
     2581 tacatcagcg ggctcccgcg gaccatgacc cagaaggacg tagaagacat gttctctcgg
     2641 tttgggcgga tcatcaactc gcgggtcctc gtggatcaga ctacaggttt gtccagaggg
     2701 gttgcgttta tccggtttga caaacggtcg gaggcagaag aggcaattac cagtttcaat
     2761 ggtcataaac ccccaggttc ctctgagccc atcacagtga agtttgcagc caaccccaac
     2821 cagaacaaaa acgtggcact cctctcgcag ctgtaccact cgccagcgcg acggttcgga
     2881 ggccccgttc accaccaggc gcagagattc aggttcgacc ccatgggcgt cgatcacatg
     2941 agcgggctct ctggcgtcaa cgtgtcagga aacgcctcct ccggctggtg cattttcatc
     3001 tacaacctgg ggcaggatgc cgacgagggg atcctctggc agatgtttgg gccgtttggt
     3061 gccgtcacca atgtgaaagt gatccgcgac ttcaacacca acaagtgcaa agggtttggc
     3121 tttgtgacca tgacaaacta tgaagaagcc gcgatggcca tagccagcct gaacggctac
     3181 cgcctggggg acaaaatctt acaggtttcc ttcaaaacca acaagtccca caaataactc
     3241 gaccctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3301 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3361 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3421 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3481 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     3541 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctta ccatgaccga
     3601 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
     3661 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
     3721 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     3781 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     3841 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     3901 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     3961 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     4021 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     4081 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     4141 gtgcccgaag gaccgcgcga cctggtgcat gacccgcaag cccggtgcct gacgcccgcc
     4201 ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catgcatcga taaaataaaa
     4261 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa
     4321 gctagcttaa gtaacgccat tttgcaaggc atggaaaaat acataactga gaatagagaa
     4381 gttcagatca aggtcaggaa cagatggaac agctgaatat gggccaaaca ggatatctgt
     4441 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggaacagctg aatatgggcc
     4501 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc
     4561 ccagatgcgg tccagccctc agcagtttct agagaaccat cagatgtttc cagggtgccc
     4621 caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt
     4681 ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcgggg
     4741 cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc
     4801 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
     4861 acccgtcagc gggggtcttt catttccgac ttgtggtctc gctgccttgg gagggtctcc
     4921 tctgagtgat tgactacccg tcagcggggg tcttcacatg cagcatgtat caaaattaat
     4981 ttggtttttt ttcttaagta tttacattaa atggccatag ttgcattaat gaatcggcca
     5041 acgcgcgggg agaggcggtt tgcgtattgg cgctcttccg cttcctcgct cactgactcg
     5101 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5161 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5221 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5281 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5341 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5401 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5461 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5521 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5581 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5641 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     5701 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5761 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5821 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5881 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5941 acctagatcc ttttaaatta aaaatgaagt ttgcggccgc aaatcaatct aaagtatata
     6001 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6061 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6121 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6181 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6241 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6301 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6361 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6421 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6481 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6541 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6601 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca
     6661 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6721 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6781 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6841 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6901 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6961 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
//
