LOCUS       Exported                9142 bp ds-DNA     circular SYN 05-ABR-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pD223-tet-pLKO.neo_shGFP
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 9142)
  AUTHORS   Krishina Ratna
  TITLE     Direct Submission
  JOURNAL   Exported quinta-feira, 5 de abr de 2018 from SnapGene 2.8.3
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9142
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     promoter        106..210
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             211..1071
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1242..1830
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2118..2139
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2154..2184
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    2192..2208
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2216..2232
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2253..2271
                     /note="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2299..2525
                     /note="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2526..2706
                     /note="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2753..2878
                     /note="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3371..3604
                     /note="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     protein_bind    4170..4188
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4198..4216
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    4223..4243
                     /note="shGFP"
     misc_feature    4250..4270
                     /note="shGFP"
     misc_feature    4317..4434
                     /note="cPPT/CTS"
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4483..4993
                     /note="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     intron          5002..5574
                     /note="beta-globin intron"
                     /note="intron from rabbit beta-globin gene"
     promoter        5629..5647
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5658..6275
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /note="TetR"
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESG"
     misc_feature    6313..6886
                     /note="IRES"
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             6893..7687
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     protein_bind    complement(7716..7815)
                     /gene="mutant version of attL"
                     /bound_moiety="LR Clonase(TM)"
                     /note="attL2"
                     /note="recombination site for the Gateway(R) LR reaction"
     LTR             7959..8192
                     /note="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8264..8385
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      8425..8560
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(8581..8599)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8609..8625)
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      8767..80
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
       61 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
      121 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
      181 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
      241 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
      301 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
      361 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
      421 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
      481 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
      541 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
      601 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
      661 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
      721 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
      781 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
      841 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
      901 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
      961 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     1021 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     1081 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     1141 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     1201 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     1261 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     1321 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     1381 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     1441 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     1501 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     1561 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     1621 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     1681 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     1741 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     1801 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     1861 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     1921 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     1981 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     2041 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     2101 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     2161 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     2221 aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag
     2281 ctggagctgc aagcttaatg tagtcttatg caatactctt gtagtcttgc aacatggtaa
     2341 cgatgagtta gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg
     2401 aagtaaggtg gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg
     2461 gacgaaccac tgaattgccg cattgcagag atattgtatt taagtgccta gctcgataca
     2521 taaacgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     2581 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     2641 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     2701 ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg
     2761 acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag
     2821 tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag
     2881 tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa
     2941 gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt
     3001 taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc
     3061 atcccttcag acaggatcag aagaacttag atcattatat aatacagtag caaccctcta
     3121 ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga
     3181 agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag
     3241 gaggagatat gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg
     3301 aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag
     3361 cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg
     3421 cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc
     3481 agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg
     3541 gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc
     3601 tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg
     3661 ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg
     3721 acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc
     3781 agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt
     3841 ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct
     3901 tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat
     3961 attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag
     4021 gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat
     4081 ctcgacggta tcgatcacga gactagcctc gagcggccgc aatatttgca tgtcgctatg
     4141 tgttctggga aatcaccata aacgtgaaat ccctatcagt gatagagact tataagttcc
     4201 ctatcagtga tagagacacc ggcaagctga ccctgaagtt catctcgaga tgaacttcag
     4261 ggtcagcttg tttttgaatt ctcgacctcg agacaaatgg cagtattcat ccacaatttt
     4321 aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca
     4381 acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt
     4441 tattacaggg acagcagaga tccactttgg ccgcggctcg agggggttgg ggttgcgcct
     4501 tttccaaggc agccctgggt ttgcgcaggg acgcggctgc tctgggcgtg gttccgggaa
     4561 acgcagcggc gccgaccctg ggtctcgcac attcttcacg tccgttcgca gcgtcacccg
     4621 gatcttcgcc gctacccttg tgggcccccc ggcgacgctt cctgctccgc ccctaagtcg
     4681 ggaaggttcc ttgcggttcg cggcgtgccg gacgtgacaa acggaagccg cacgtctcac
     4741 tagtaccctc gcagacggac agcgccaggg agcaatggca gcgcgccgac cgcgatgggc
     4801 tgtggccaat agcggctgct cagcagggcg cgccgagagc agcggccggg aaggggcggt
     4861 gcgggaggcg gggtgtgggg cggtagtgtg ggccctgttc ctgcccgcgc ggtgttccgc
     4921 attctgcaag cctccggagc gcacgtcggc agtcggctcc ctcgttgacc gaatcaccga
     4981 cctctctccc cagggggatc tgtgagtttg gggacccttg attgttcttt ctttttcgct
     5041 attgtaaaat tcatgttata tggagggggc aaagttttca gggtgttgtt tagaatggga
     5101 agatgtccct tgtatcacca tggaccctca tgataatttt gtttctttca ctttctactc
     5161 tgttgacaac cattgtctcc tcttattttc ttttcatttt ctgtaacttt ttcgttaaac
     5221 tttagcttgc atttgtaacg aatttttaaa ttcacttttg tttatttgtc agattgtaag
     5281 tactttctct aatcactttt ttttcaaggc aatcagggta tattatattg tacttcagca
     5341 cagttttaga gaacaattgt tataattaaa tgataaggta gaatatttct gcatataaat
     5401 tctggctggc gtggaaatat tcttattggt agaaacaact acatcctggt catcatcctg
     5461 cctttctctt tatggttaca atgatataca ctgtttgaga tgaggataaa atactctgag
     5521 tccaaaccgg gcccctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     5581 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattgta atacgactca
     5641 ctatagggcg aattgatatg tctagattag ataaaagtaa agtgattaac agcgcattag
     5701 agctgcttaa tgaggtcgga atcgaaggtt taacaacccg taaactcgcc cagaagctag
     5761 gtgtagagca gcctacattg tattggcatg taaaaaataa gcgggctttg ctcgacgcct
     5821 tagccattga gatgttagat aggcaccata ctcacttttg ccctttagaa ggggaaagct
     5881 ggcaagattt tttacgtaat aacgctaaaa gttttagatg tgctttacta agtcatcgcg
     5941 atggagcaaa agtacattta ggtacacggc ctacagaaaa acagtatgaa actctcgaaa
     6001 atcaattagc ctttttatgc caacaaggtt tttcactaga gaatgcatta tatgcactca
     6061 gcgctgtggg gcattttact ttaggttgcg tattggaaga tcaagagcat caagtcgcta
     6121 aagaagaaag ggaaacacct actactgata gtatgccgcc attattacga caagctatcg
     6181 aattatttga tcaccaaggt gcagagccag ccttcttatt cggccttgaa ttgatcatat
     6241 gcggattaga aaaacaactt aaatgtgaaa gtgggtccgc gtacagcggc tcccgggagt
     6301 tctagggatc tgcccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga
     6361 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa
     6421 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
     6481 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc
     6541 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
     6601 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca
     6661 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag
     6721 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct
     6781 ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc
     6841 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataaggat ccatgattga
     6901 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     6961 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     7021 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     7081 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     7141 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     7201 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     7261 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     7321 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     7381 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     7441 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     7501 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     7561 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     7621 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     7681 cttctgaggt acctttttta agggtgggcg cgccgaccca gctttcttgt acaaagttgg
     7741 cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
     7801 taaaatcatt atttgccatc cagctgatat ccgcccgccc cacgacccgc agcgcccgac
     7861 cgaaaggagc gcacgacccc atgcatcggt acctttaaga ccaatgactt acaaggcagc
     7921 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
     7981 acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc
     8041 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     8101 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     8161 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca
     8221 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc
     8281 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     8341 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta
     8401 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     8461 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     8521 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta cccaattcgc
     8581 cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg
     8641 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     8701 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     8761 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     8821 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     8881 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     8941 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     9001 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     9061 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     9121 atttataagg gattttgccg at
//
