LOCUS       sequence-146916-        8008 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  tet-inducible human KLF4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8008)
  AUTHORS   Takashima Y, Guo G, Loos R, Nichols J, Ficz G, Krueger F, Oxley D, 
            Santos F, Clarke J, Mansfield W, Reik W, Bertone P, Smith A
  TITLE     Resetting Transcription Factor Control Circuitry toward Ground-State
            Pluripotency in Human.
  JOURNAL   Cell. 2014 Sep 11;158(6):1254-69. doi: 10.1016/j.cell.2014.08.029.
  PUBMED    25215486
REFERENCE   2  (bases 1 to 8008)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2014.08"; journalName: "Cell"; date: "2014-09-11- 
            11"; volume: "158"; issue: "6"; pages: "1254-69"
COMMENT     Sequence Label: pPB-hCMV1-hKLF2-IV
FEATURES             Location/Qualifiers
     source          1..8008
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     623..642
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   660..694
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    join(1075..1093,1094..1116,1117..1135,1136..1158,1159..1177,
                     1178..1200,1201..1219,1220..1242,1243..1261,1262..1284,
                     1285..1303,1304..1326,1327..1345)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        1378..1416
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1413..1437
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          1539..2111
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     protein_bind    2176..2200
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3302..3326)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3364..3390
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    join(3426..3999,4000..4002,4003..4011)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3593..3610)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3820..3839
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4012..4014,4015..4017,4018..4731)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4057..4078)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4318..4337)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4665..4686
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5116..5133)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5122..5346
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5393..5409)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5417..5433
                     /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(join(5441..5447,5448..5465,5466..5471))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5486..5507
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   5734..5796
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(5820..5838)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5859..5875)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5883..5899
                     /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(join(5907..5913,5914..5931,5932..5937))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5952..5973
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6090..6107)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6261..6849)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6341..6360)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7020..7811,7812..7880))
                     /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     7643..7662
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7881..7985)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg gtaccttaac
      661 cctagaaaga tagtctgcgt aaaattgacg catgcattct tgaaatattg ctctctcttt
      721 ctaaatagcg cgaatccgtc gctgtgcatt taggacatct cagtcgccgc ttggagctcc
      781 cgtgaggcgt gcttgtcaat gcggtaagtg tcactgattt tgaactataa cgaccgcgtg
      841 agtcaaaatg acgcatgatt atcttttacg tgacttttaa gatttaactc atacgataat
      901 tatattgtta tttcatgttc tacttacgtg ataacttatt atatatatat tttcttgtta
      961 tagatatcat aatttaaaca agcaaaacca aattaagggc cagctcattc ctcccactca
     1021 tgatctatag atctggatcc gtcgacaacg gcaagcttac atcgagttta ccactcccta
     1081 tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt
     1141 gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac
     1201 tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag
     1261 aaaagtgaaa gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt
     1321 taccactccc tatcagtgat agagaaaagt gaaagtcgag ctcggtaccc gggtcgaggt
     1381 aggcgtgtac ggtgggaggc ctatataagc agagctcgtt tagtgaaccg tcagatcgcc
     1441 tggagacgcc atccacgctg ttttgacctc catagaagac accgggaccg atccagcctc
     1501 cgcgggactc agacacccgg gatcctgaga acttcagggt gagtttgggg acccttgatt
     1561 gttctttctt tttcgctatt gtaaaattca tgttatatgg agggggcaaa gttttcaggg
     1621 tgttgtttag aatgggaaga tgtcccttgt atcaccatgg accctcatga taattttgtt
     1681 tctttcactt tctactctgt tgacaaccat tgtctcctct tattttcttt tcattttctg
     1741 taactttttc gttaaacttt agcttgcatt tgtaacgaat ttttaaattc acttttgttt
     1801 atttgtcaga ttgtaagtac tttctctaat cacttttttt tcaaggcaat cagggtatat
     1861 tatattgtac ttcagcacag ttttagagaa caattgttat aattaaatga taaggtagaa
     1921 tatttctgca tataaattct ggctggcgtg gaaatattct tattggtaga aacaactaca
     1981 ccctggtcat catcctgcct ttctctttat ggttacaatg atatacactg tttgagatga
     2041 ggataaaata ctctgagtcc aaaccgggcc cctctgctaa ccatgttcat gccttcttct
     2101 ctttcctaca gctcctgggc aacgtgctgg ttgttgtgct gtctcatcat tttggcaaag
     2161 aattcctcga atcaaacaag tttgtacaaa aaagcaggct tcgaaggaga tagaaccatg
     2221 gcgctgagtg aacccatcct gccgtccttc tccactttcg ccagcccgtg ccgcgagcgc
     2281 ggcctgcagg agcgctggcc gcgcgccgaa cccgagtccg gcggcaccga cgacgacctc
     2341 aacagcgtgc tggacttcat cctgtccatg gggctggatg gcctgggcgc cgaggccgcc
     2401 ccggagccgc cgccgccgcc cccgccgcct gcgttctatt accccgaacc cggcgcgccc
     2461 ccgccctaca gcgcccccgc gggtggcctg gtgtctgagc tgctgcgacc cgagctggat
     2521 gcgccgccgg ggcccgcact gcacggccgc tttctgctgg cgccgcccgg ccgcctggtc
     2581 aaggccgagc cccctgaagc ggacggcggc ggcggctacg gctgcgcccc cgggctgacc
     2641 cgtggaccgc gcggcctcaa gcgcgagggc gccccgggcc cggcggcttc gtgcatgcga
     2701 ggtcccgggg gccgcccccc gccgccgccc gacacaccgc cgctcagccc cgacggcccc
     2761 gcgcgcctgc ccgcgcccgg tccgcgcgcc tccttcccgc cgcctttcgg tggccctggt
     2821 ttcggcgcgc ccgggcccgg cctgcattac gcgccgcctg cgcccccagc cttcggtctc
     2881 ttcgacgacg cggccgccgc cgcggcagcc ctgggcctgg cgccccccgc cgcccgcggt
     2941 ctcctcacgc cgcctgcgtc cccgctggag ctgctggagg ccaagccaaa gcgcggccgc
     3001 cgctcttggc cccgcaaacg caccgccact cacacctgca gctacgcggg ctgcggcaag
     3061 acctacacca agagttcgca tctgaaggcg catctgcgca cgcacacagg tgagaagccc
     3121 taccactgca actgggacgg ctgcggctgg aagtttgcgc gctcagacga gctcacgcgc
     3181 cactaccgaa agcacacggg ccaccggcca ttccagtgcc atctgtgcga tcgtgccttc
     3241 tcgcgctccg atcacctggc gctgcacatg aaacggcaca tgtagggatc cnacccagct
     3301 tacccagctt tcttgtacaa agtggttcga ttcgagatct cacgcgtgac tagtgccatg
     3361 gcctacccat acgacgtgcc agactacgcc tagcggccgc tcgataagct tgatatcgaa
     3421 ttccgcccct ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     3481 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     3541 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     3601 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
     3661 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     3721 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     3781 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     3841 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     3901 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa acgtctaggc cccccgaacc
     3961 acggggacgt ggttttcctt tgaaaaacac gatgataata tggccacaac catggtgagc
     4021 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     4081 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     4141 accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     4201 accctgggct acggcctgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
     4261 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     4321 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     4381 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     4441 tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag
     4501 gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac
     4561 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     4621 taccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     4681 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta aagcggccag
     4741 cttatcgact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat
     4801 agtcgagaaa ttgatgatct attaaacaat aaagatgtcc acatggaagt ttttcctgtc
     4861 atactttgtt aagaagggtg agaacagagt acctacattt tgaatggaag gattggagct
     4921 acgggggtgg gggtggggtg ggattagata aatgcctgct ctttactgaa ggctctttac
     4981 tattgcttta tgataatgtt tcatagttgg atatcataat ttaaacaagc aaaaccaaat
     5041 taagggccag ctcattcctc ccactcatga tctatagatc ctaagcttat cgatgatcta
     5101 gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     5161 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     5221 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     5281 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     5341 ctatggcttc tgaggcggaa agaacctgca gcccaagctt ggcgtaatca tggtcatagc
     5401 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5461 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5521 cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatcctttgt tactttatag
     5581 aagaaatttt gagtttttgt ttttttttaa taaataaata aacataaata aattgtttgt
     5641 tgaatttatt attagtatgt aagtgtaaat ataataaaac ttaatatcta ttcaaattaa
     5701 taaataaacc tcgatataca gaccgataaa acacatgcgt caattttacg catgattatc
     5761 tttaacgtac gtcacaatat gattatcttt ctagggttaa agagctccag cttttgttcc
     5821 ctttagtgag ggttaatttc gagcttggcg taatcatggt catagctgtt tcctgtgtga
     5881 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     5941 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     6001 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     6061 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     6121 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     6181 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6241 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6301 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6361 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6421 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     6481 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6541 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6601 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6661 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     6721 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6781 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6841 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6901 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6961 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     7021 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     7081 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     7141 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     7201 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7261 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7321 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7381 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7441 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7501 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7561 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7621 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7681 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7741 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7801 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7861 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7921 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7981 ccgcgcacat ttccccgaaa agtgccac
//
