LOCUS       FUW-tetO-hKLF4.        9904 bp DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    FUW-tetO-hKLF4.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9904)
  AUTHORS   Hockemeyer D, Soldner F, Cook EG, Gao Q, Mitalipova M, Jaenisch R
  TITLE     A drug-inducible system for direct reprogramming of human somatic 
            cells to pluripotency.
  JOURNAL   Cell Stem Cell. 2008 Sep 11. 3(3):346-53.
  PUBMED    18786421
REFERENCE   2  (bases 1 to 9904)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 9904)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Stem 
            Cell. 2008 Sep 11. 3(3):346-53."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9904
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        7..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..589
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     LTR             604..784
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    831..956
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1449..1682
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1867..1911
                     /label=gp41 peptide
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et 
                     al., 2013)"
     CDS             2060..2101
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     misc_feature    2209..2326
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     protein_bind    2385..2655
                     /label=tetracycline response element
                     /note="contains seven copies of the tetracycline operator
                     tetO"
     promoter        2688..2725
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     primer_bind     2722..2746
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2901..4337
                     /label=hKLF4
                     /note="Homo sapiens Kruppel-like factor 4 (Klf4) gene.
                     Belongs to the relatively large family of SP1-like 
                     transcription factors and is involved in the regulation of 
                     proliferation, differentiation, apoptosis and somatic cell 
                     reprogramming."
     misc_feature    4365..4953
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     primer_bind     complement(4956..4972)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             5478..5658
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    5690..5914
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5960..6388
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6402..6731
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        6779..6826
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             6845..7216
                     /label=BleoR
                     /note="antibiotic-binding protein"
     polyA_signal    7349..7482
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7519..7535)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(7543..7559)
                     /label=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)."
     promoter        complement(7567..7597)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7612..7633)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     primer_bind     complement(7750..7767)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7921..8509)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8683..9540)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(9541..9645)
                     /label=AmpR promoter
     primer_bind     complement(9720..9739)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
ORIGIN
        1 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt
      601 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
      661 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
      721 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
      781 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
      841 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
      901 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
      961 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
     1021 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
     1081 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
     1141 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
     1201 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
     1261 agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga
     1321 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
     1381 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
     1441 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
     1501 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
     1561 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
     1621 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
     1681 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
     1741 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
     1801 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
     1861 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
     1921 tttaacataa caaattggct gtggtatata aaattattca taatgatagt aggaggcttg
     1981 gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat
     2041 tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga
     2101 atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatcg
     2161 gcactgcgtg cgccaattct gcagacaaat ggcagtattc atccacaatt ttaaaagaaa
     2221 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
     2281 acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag
     2341 ggacagcaga gatccagttt ggttaatgat ctcgagttta ccactcccta tcagtgatag
     2401 agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag
     2461 tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag
     2521 tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa
     2581 gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc
     2641 tatcagtgat agagaaaagt gaaagtcgag ctcggtaccc gggtcgagta ggcgtgtacg
     2701 gtgggaggcc tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca
     2761 tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc gcggccccga
     2821 attccttctc ttcgttgact ttggggttca ggtgccccag ctgcttcggg ctgccgagga
     2881 ccttctgggc ccccacatta atgaggcagc cacctggcga gtctgacatg gctgtcagcg
     2941 acgcgctgct cccatctttc tccacgttcg cgtctggccc ggcgggaagg gagaagacac
     3001 tgcgtcaagc aggtgccccg aataaccgct ggcgggagga gctctcccac atgaagcgac
     3061 ttcccccagt gcttcccggc cgcccctatg acctggcggc ggcgaccgtg gccacagacc
     3121 tggagagcgg cggagccggt gcggcttgcg gcggtagcaa cctggcgccc ctacctcgga
     3181 gagagaccga ggagttcaac gatctcctgg acctggactt tattctctcc aattcgctga
     3241 cccatcctcc ggagtcagtg gccgccaccg tgtcctcgtc agcgtcagcc tcctcttcgt
     3301 cgtcgccgtc gagcagcggc cctgccagcg cgccctccac ctgcagcttc acctatccga
     3361 tccgggccgg gaacgacccg ggcgtggcgc cgggcggcac gggcggaggc ctcctctatg
     3421 gcagggagtc cgctccccct ccgacggctc ccttcaacct ggcggacatc aacgacgtga
     3481 gcccctcggg cggcttcgtg gccgagctcc tgcggccaga attggacccg gtgtacattc
     3541 cgccgcagca gccgcagccg ccaggtggcg ggctgatggg caagttcgtg ctgaaggcgt
     3601 cgctgagcgc ccctggcagc gagtacggca gcccgtcggt catcagcgtc agcaaaggca
     3661 gccctgacgg cagccacccg gtggtggtgg cgccctacaa cggcgggccg ccgcgcacgt
     3721 gccccaagat caagcaggag gcggtctctt cgtgcaccca cttgggcgct ggaccccctc
     3781 tcagcaatgg ccaccggccg gctgcacacg acttccccct ggggcggcag ctccccagca
     3841 ggactacccc gaccctgggt cttgaggaag tgctgagcag cagggactgt caccctgccc
     3901 tgccgcttcc tcccggcttc catccccacc cggggcccaa ttacccatcc ttcctgcccg
     3961 atcagatgca gccgcaagtc ccgccgctcc attaccaaga gctcatgcca cccggttcct
     4021 gcatgccaga ggagcccaag ccaaagaggg gaagacgatc gtggccccgg aaaaggaccg
     4081 ccacccacac ttgtgattac gcgggctgcg gcaaaaccta cacaaagagt tcccatctca
     4141 aggcacacct gcgaacccac acaggtgaga aaccttacca ctgtgactgg gacggctgtg
     4201 gatggaaatt cgcccgctca gatgaactga ccaggcacta ccgtaaacac acggggcacc
     4261 gcccgttcca gtgccaaaaa tgcgaccgag cattttccag gtcggaccac ctcgccttac
     4321 acatgaagag gcatttttaa gaattcgata tcaagcttat cgataatcaa cctctggatt
     4381 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     4441 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     4501 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     4561 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     4621 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     4681 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     4741 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
     4801 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     4861 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     4921 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgacct cgagacctag
     4981 aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc
     5041 tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac
     5101 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     5161 aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac
     5221 aaggctactt ccctgattgg cagaactaca caccagggcc agggatcaga tatccactga
     5281 cctttggatg gtgctacaag ctagtaccag ttgagcaaga gaaggtagaa gaagccaatg
     5341 aaggagagaa cacccgcttg ttacaccctg tgagcctgca tgggatggat gacccggaga
     5401 gagaagtatt agagtggagg tttgacagcc gcctagcatt tcatcacatg gcccgagagc
     5461 tgcatccgga ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     5521 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     5581 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     5641 tgtggaaaat ctctagcagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     5701 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     5761 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     5821 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     5881 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     5941 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     6001 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     6061 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     6121 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     6181 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     6241 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     6301 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     6361 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     6421 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     6481 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     6541 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     6601 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     6661 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     6721 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt
     6781 tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta
     6841 aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc
     6901 ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc
     6961 cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc
     7021 ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc
     7081 ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga
     7141 gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt
     7201 ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg ccttctatga
     7261 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     7321 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     7381 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     7441 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     7501 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     7561 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     7621 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     7681 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     7741 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     7801 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     7861 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     7921 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     7981 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     8041 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     8101 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     8161 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     8221 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     8281 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     8341 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     8401 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     8461 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     8521 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     8581 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     8641 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     8701 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     8761 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     8821 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     8881 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     8941 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     9001 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     9061 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     9121 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     9181 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     9241 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     9301 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     9361 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     9421 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     9481 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     9541 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     9601 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     9661 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     9721 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     9781 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     9841 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
     9901 tata
//
