LOCUS       Exported                8545 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  A lentiviral vector for the expression of mouse Olig1 under 
            doxycyline control.
ACCESSION   .
VERSION     .
KEYWORDS    pLVX-Tight-Puro_Olig1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8545)
  AUTHORS   Najm FJ, Lager AM, Zaremba A, Wyatt K, Caprariello AV, Factor DC, 
            Karl RT, Maeda T, Miller RH, Tesar PJ
  TITLE     Transcription factor-mediated reprogramming of fibroblasts to 
            expandable, myelinogenic oligodendrocyte progenitor cells.
  JOURNAL   Nat Biotechnol. 2013 May;31(5):426-33. doi: 10.1038/nbt.2561. Epub 
            2013 Apr 14.
  PUBMED    23584611
REFERENCE   2  (bases 1 to 8545)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8545
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2028..2143
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        2206..2520
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    2214..2232
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2250..2268
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2285..2303
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2321..2339
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2357..2375
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2392..2410
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2428..2446
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2493..2517
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        3330..3829
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3365..3388)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3762..3781
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3850..4449
                     /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(3850..3869)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4346..4366
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4467..5055
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4520..4540)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4938..4949)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5263..5896
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6025..6041)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6025..6041)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6038..6060)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6049..6065
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     promoter        complement(6073..6103)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6118..6139
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6256..6273)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6427..7012)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6507..6526)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7183..8043)
                     /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     7806..7825
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8044..8148)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    8196..8330
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8233..8252)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8287..8306
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga actacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgatgaggcc ctttcgtctt cactcgagtt tactccctat
     2221 cagtgataga gaacgtatgt cgagtttact ccctatcagt gatagagaac gatgtcgagt
     2281 ttactcccta tcagtgatag agaacgtatg tcgagtttac tccctatcag tgatagagaa
     2341 cgtatgtcga gtttactccc tatcagtgat agagaacgta tgtcgagttt atccctatca
     2401 gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacgt atgtcgaggt
     2461 aggcgtgtac ggtgggaggc ctatataagc agagctcgtt tagtgaaccg tcagatcgcc
     2521 tggagaagga tccaccatgt actatgcgat ttcccaggcg cgtgtgaacg cggctcccgc
     2581 gaccatgttg cggccacagc ggcctggaga cgtgcagctc ggggcctcgc tgtatgagct
     2641 ggtgggttac aggcagccac ctatctcctc atcctcatcc tcatcctctt ccaccgcatc
     2701 ccttctcccc aagcctgcgc gcgagaaggc cgaagctccg ctcgctgaac ctcggggacc
     2761 tgcgccagag tccgggggcg cccgggccga cgccaaagag gaacagcagc agcagcagct
     2821 gcggcgcaag atcaacagcc gcgagcggaa gcgcatgcag gacctgaact tggccatgga
     2881 cgcgctgcgc gaagttatcc taccctactc ggcagcgcac tgccagggcg cgccgggccg
     2941 caagctctcc aagattgcca cgctgctgct cgcccgcaac tacatcctgc tgctgggtag
     3001 ctcgctgcag gagctgcgcc gcgcgctcgg cgacggtgcg ggacccgccg ccccgcgcct
     3061 gctcctcgct ggcctgcctc ttctggctgc cgcgccgggc tctgtgctgc tggcacccgg
     3121 tgccgtggga ccccccgaaa cgctgcgccc caccaagtac ctgtctctag cgctcgacga
     3181 gccaccgtgc ggccagttct ctctcccagc tggtggcgcg ggtagccctg gcctctgctc
     3241 ctgtgcggtg tgcaagttcc cgcatctggt ccccgccggc ctgggcttgg cagcggtgca
     3301 ggcccagttc tccaagtagg aattctaccg ggtaggggag gcgcttttcc caaggcagtc
     3361 tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg gcctctggcc
     3421 tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt
     3481 cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc
     3541 gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac
     3601 cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt
     3661 cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct
     3721 caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa
     3781 gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga cctgcagccc
     3841 aagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     3901 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     3961 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     4021 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     4081 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     4141 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     4201 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     4261 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     4321 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     4381 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     4441 ggtgcctgac ggggcgcgtc tggaacaatc aacctctgga ttacaaaatt tgtgaaagat
     4501 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     4561 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     4621 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     4681 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     4741 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     4801 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     4861 agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     4921 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     4981 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     5041 gggccgcctc cccgcctgga attaattctg cagtcgagac ctagaaaaac atggagcaat
     5101 cacaagtagc aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga
     5161 ggaggaggtg ggtttttcca gtcacacctc aggtaccttt aagaccaatg acttacaagg
     5221 cagctgtaga tcttagccac tttttaaaag aaaagagggg actggaaggg ctaattcact
     5281 cccaacgaag acaagatatc cttgatctgt ggatctacca cacacaaggc tacttccctg
     5341 attagcagaa ctacacacca gggccagggg tcagatatcc actgaccttt ggatggtgct
     5401 acaagctagt accagttgag ccagataagg tagaagaggc caataaagga gagaacacca
     5461 gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa gtgttagagt
     5521 ggaggtttga cagccgccta gcatttcatc acgtggcccg agagctgcat ccggagtact
     5581 tcaagaactg ctgatatcga gcttgctaca agggactttc cgctggggac tttccaggga
     5641 ggcgtggcct gggcgggact ggggagtggc gagccctcag atcctgcata taagcagctg
     5701 ctttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     5761 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     5821 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     5881 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca
     5941 aagaaatgaa tatcagagag tgagaggcct tgacattgct agcgttttac cgtcgacctc
     6001 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     6061 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     6121 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6181 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6241 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6301 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6361 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6421 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6481 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6541 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6601 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6661 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6721 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6781 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6841 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6901 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6961 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7021 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7081 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7141 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7201 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7261 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7321 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7381 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7441 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7501 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7561 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7621 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7681 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7741 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7801 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7861 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7921 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7981 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8041 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8101 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8161 aaaagtgcca cctgacgtcg acggatcggg agatcaactt gtttattgca gcttataatg
     8221 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     8281 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc aactggataa
     8341 ctcaagctaa ccaaaatcat cccaaacttc ccaccccata ccctattacc actgccaatt
     8401 acctagtggt ttcatttact ctaaacctgt gattcctctg aattattttc attttaaaga
     8461 aattgtattt gttaaatatg tactacaaac ttagtagttt ttaaagaaat tgtatttgtt
     8521 aaatatgtac tacaaactta gtagt
//
