LOCUS       Exported                7681 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Dox-inducible lentiviral vector expressing mouse Klf4.
ACCESSION   .
VERSION     .
KEYWORDS    pHAGE2-TetOminiCMV-Klf4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7681)
  AUTHORS   Velychko S, Adachi K, Kim KP, Hou Y, MacCarthy CM, Wu G, Scholer HR
  TITLE     Excluding Oct4 from Yamanaka Cocktail Unleashes the Developmental 
            Potential of iPSCs.
  JOURNAL   Cell Stem Cell. 2019 Oct 30. pii: S1934-5909(19)30423-0. doi: 
            10.1016/j.stem.2019.10.002.
  PUBMED    31708402
REFERENCE   2  (bases 1 to 7681)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7681
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     56..80
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      146..155
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    1590..2178
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1643..1663)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2061..2072)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2326..2506
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(2553..2571)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2639..2743
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2744..3604
                     /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     complement(2962..2981)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3775..4363
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4264..4283
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4517..4534
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4626..4645
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     protein_bind    4774..4795
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4810..4840
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4848..4864
                     /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)."
     primer_bind     4853..4875
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4872..4888
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4872..4888
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4938..4957
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             5149..5782
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    5829..5954
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6451..6684
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6869..6913
                     /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    7206..7323
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    7399..7669
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
ORIGIN
        1 gagctcggta cccgggtcga ggtaggcatg tacggtggga ggcctatata agcagagctc
       61 gtttagtgaa ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa
      121 gacaccggga ccgatccagc ctgcggccgc catggctgtc agcgacgctc tgctcccgtc
      181 cttctccacg ttcgcgtccg gcccggcggg aagggagaag acactgcgtc cagcaggtgc
      241 cccgactaac cgttggcgtg aggaactctc tcacatgaag cgacttcccc cacttcccgg
      301 ccgcccctac gacctggcgg cgacggtggc cacagacctg gagagtggcg gagctggtgc
      361 agcttgcagc agtaacaacc cggccctcct agcccggagg gagaccgagg agttcaacga
      421 cctcctggac ctagacttta tcctttccaa ctcgctaacc caccaggaat cggtggccgc
      481 caccgtgacc acctcggcgt cagcttcatc ctcgtcttcc ccagcgagca gcggccctgc
      541 cagcgcgccc tccacctgca gcttcagcta tccgatccgg gccgggggtg acccgggcgt
      601 ggctgccagc aacacaggtg gagggctcct ctacagccga gaatctgcgc cacctcccac
      661 ggcccccttc aacctggcgg acatcaatga cgtgagcccc tcgggcggct tcgtggctga
      721 gctcctgcgg ccggagttgg acccagtata cattccgcca cagcagcctc agccgccagg
      781 tggcgggctg atgggcaagt ttgtgctgaa ggcgtctctg accacccctg gcagcgagta
      841 cagcagccct tcggtcatca gtgttagcaa aggaagccca gacggcagcc accccgtggt
      901 agtggcgccc tacagcggtg gcccgccgcg catgtgcccc aagattaagc aagaggcggt
      961 cccgtcctgc acggtcagcc ggtccctaga ggcccatttg agcgctggac cccagctcag
     1021 caacggccac cggcccaaca cacacgactt ccccctgggg cggcagctcc ccaccaggac
     1081 tacccctaca ctgagtcccg aggaactgct gaacagcagg gactgtcacc ctggcctgcc
     1141 tcttccccca ggattccatc cccatccggg gcccaactac cctcctttcc tgccagacca
     1201 gatgcagtca caagtcccct ctctccatta tcaagagctc atgccaccgg gttcctgcct
     1261 gccagaggag cccaagccaa agaggggaag aaggtcgtgg ccccggaaaa gaacagccac
     1321 ccacacttgt gactatgcag gctgtggcaa aacctatacc aagagttctc atctcaaggc
     1381 acacctgcga actcacacag gcgagaaacc ttaccactgt gactgggacg gctgtgggtg
     1441 gaaattcgcc cgctcggatg aactgaccag gcactaccgc aaacacacag ggcaccggcc
     1501 ctttcagtgc cagaagtgcg acagggcctt ttccaggtcg gaccaccttg ccttacacat
     1561 gaagaggcac ttttaaatcg atagatccta atcaacctct ggattacaaa atttgtgaaa
     1621 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     1681 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     1741 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     1801 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     1861 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     1921 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     1981 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     2041 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     2101 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     2161 tttgggccgc ctccccgcct ggtaccttta agaccaatga cttacaaggc agctgtagat
     2221 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
     2281 caagatcacc tgcaggacag gcgcgccctg ctttttgctt gtactgggtc tctctggtta
     2341 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     2401 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     2461 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcacccg ggcgattaag
     2521 gaaagggcta gatcattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     2581 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     2641 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     2701 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     2761 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     2821 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     2881 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     2941 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     3001 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     3061 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     3121 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     3181 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     3241 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     3301 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     3361 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     3421 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     3481 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     3541 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     3601 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     3661 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     3721 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3781 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3841 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3901 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3961 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4021 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4081 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4141 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4201 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4261 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4321 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4381 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     4441 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     4501 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     4561 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc aagctcatgg ctgactaatt
     4621 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     4681 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc ccgtggcacg acaggtttcc
     4741 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     4801 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     4861 acaatttcac acaggaaaca gctatgacat gattacgaat ttcacaaata aagcattttt
     4921 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     4981 caactggata actcaagcta accaaaatca tcccaaactt cccaccccat accctattac
     5041 cactgccaat tacctgtggt ttcatttact ctaaacctgt gattcctctg aattattttc
     5101 attttaaaga aattgtattt gttaaatatg tactacaaac ttagtagttg gaagggctaa
     5161 ttcactccca aagaagacaa gatatccttg atctgtggat ctaccacaca caaggctact
     5221 tccctgatta gcagaactac acaccagggc caggggtcag atatccactg acctttggat
     5281 ggtgctacaa gctagtacca gttgagccag ataaggtaga agaggccaat aaaggagaga
     5341 acaccagctt gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtgt
     5401 tagagtggag gtttgacagc cgcctagcat ttcatcacgt ggcccgagag ctgcatccgg
     5461 agtacttcaa gaactgctga tatcgagctt gctacaaggg actttccgct ggggactttc
     5521 cagggaggcg tggcctgggc gggactgggg agtggcgagc cctcagatcc tgcatataag
     5581 cagctgcttt ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     5641 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     5701 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     5761 tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac
     5821 cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg
     5881 ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg
     5941 ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt
     6001 taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc
     6061 tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac
     6121 tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata
     6181 cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt
     6241 tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccggcc
     6301 gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg aattatataa
     6361 atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt
     6421 ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc
     6481 agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt
     6541 gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct
     6601 gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag
     6661 atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac
     6721 cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca
     6781 cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt
     6841 aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa
     6901 atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt
     6961 cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt
     7021 gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag
     7081 gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc
     7141 cattcgatta gtgaacggat ctcgacggta tcgccgaatt cacaaatggc agtattcatc
     7201 cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac
     7261 ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat
     7321 tttcgggttt attacaggga cagcagagat ccagtttgga ctagtccaca ccctaactga
     7381 cacactcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac
     7441 tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag
     7501 aaaagtgaaa gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt
     7561 taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg
     7621 atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt
     7681 c
//
