LOCUS       sequence-410034-        7715 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human KLF4 in mammalian cells, for lentivirus generation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7715)
  AUTHORS   Weng M, Hu H, Graus MS, Tan DS, Gao Y, Ren S, Ho DHH, Langer J, 
            Holzner M, Huang Y, Ling GS, Lai CSW, Francois M, Jauch R
  TITLE     An engineered Sox17 induces somatic to neural stem cell fate 
            transitions independently from pluripotency reprogramming.
  JOURNAL   Sci Adv. 2023 Aug 25;9(34):eadh2501. doi: 10.1126/sciadv.adh2501. 
            Epub 2023 Aug 23.
  PUBMED    37611093
REFERENCE   2  (bases 1 to 7715)
  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"; journalName: "Sci Adv."; 
            date: "2023-08-25"; volume: "9(34):eadh2501. doi"; pages: " 
            10.1126/sciadv.adh2501. Epub 2023 Aug 23"
COMMENT     Sequence Label: pHAGE2-TetO-hKLF4
FEATURES             Location/Qualifiers
     source          1..7715
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    join(2251..2269,2270..2292,2293..2311,2312..2334,2335..2353,
                     2354..2376,2377..2395,2396..2418,2419..2437,2438..2460,
                     2461..2479,2480..2502,2503..2521)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     primer_bind     2589..2613
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     misc_feature    4123..4711
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4176..4196)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4594..4605)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(4841..4874)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             4893..5073
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5120..5138)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5206..5310
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5311..5379,5380..6171)
                     /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(5529..5548)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6342..6930
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7084..7101
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7341..7362
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7377..7382,7383..7400,7401..7407)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7415..7431
                     /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     7420..7442
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7439..7455
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7439..7455
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7505..7524
                     /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 tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtcca
     2221 caccctaact gacacactcg agtttaccac tccctatcag tgatagagaa aagtgaaagt
     2281 cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta
     2341 tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt
     2401 gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac
     2461 tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag
     2521 aaaagtgaaa gtcgagctcg gtacccgggt cgaggtaggc atgtacggtg ggaggcctat
     2581 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
     2641 gacctccata gaagacaccg ggaccgatcc agcctgcggc caccaccggt atggctgtca
     2701 gcgacgcgct gctcccatct ttctccacgt tcgcgtctgg cccggcggga agggagaaga
     2761 cactgcgtca agcaggtgcc ccgaataacc gctggcggga ggagctctcc cacatgaagc
     2821 gacttccccc agtgcttccc ggccgcccct atgacctggc ggcggcgacc gtggccacag
     2881 acctggagag cggcggagcc ggtgcggctt gcggcggtag caacctggcg cccctacctc
     2941 ggagagagac cgaggagttc aacgatctcc tggacctgga ctttattctc tccaattcgc
     3001 tgacccatcc tccggagtca gtggccgcca ccgtgtcctc gtcagcgtca gcctcctctt
     3061 cgtcgtcgcc gtcgagcagc ggccctgcca gcgcgccctc cacctgcagc ttcacctatc
     3121 cgatccgggc cgggaacgac ccgggcgtgg cgccgggcgg cacgggcgga ggcctcctct
     3181 atggcaggga gtccgctccc cctccgacgg ctcccttcaa cctggcggac atcaacgacg
     3241 tgagcccctc gggcggcttc gtggccgagc tcctgcggcc agaattggac ccggtgtaca
     3301 ttccgccgca gcagccgcag ccgccaggtg gcgggctgat gggcaagttc gtgctgaagg
     3361 cgtcgctgag cgcccctggc agcgagtacg gcagcccgtc ggtcatcagc gtcagcaaag
     3421 gcagccctga cggcagccac ccggtggtgg tggcgcccta caacggcggg ccgccgcgca
     3481 cgtgccccaa gatcaagcag gaggcggtct cttcgtgcac ccacttgggc gctggacccc
     3541 ctctcagcaa tggccaccgg ccggctgcac acgacttccc cctggggcgg cagctcccca
     3601 gcaggactac cccgaccctg ggtcttgagg aagtgctgag cagcagggac tgtcaccctg
     3661 ccctgccgct tcctcccggc ttccatcccc acccggggcc caattaccca tccttcctgc
     3721 ccgatcagat gcagccgcaa gtcccgccgc tccattacca agagctcatg ccacccggtt
     3781 cctgcatgcc agaggagccc aagccaaaga ggggaagacg atcgtggccc cggaaaagga
     3841 ccgccaccca cacttgtgat tacgcgggct gcggcaaaac ctacacaaag agttcccatc
     3901 tcaaggcaca cctgcgaacc cacacaggtg agaaacctta ccactgtgac tgggacggct
     3961 gtggatggaa attcgcccgc tcagatgaac tgaccaggca ctaccgtaaa cacacggggc
     4021 accgcccgtt ccagtgccaa aaatgcgacc gagcattttc caggtcggac cacctcgcct
     4081 tacacatgaa gaggcatttt taagtcgaca tcgatagatc ctaatcaacc tctggattac
     4141 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     4201 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     4261 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     4321 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     4381 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     4441 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     4501 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     4561 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     4621 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     4681 agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa tgacttacaa
     4741 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     4801 ctcccaacga agacaagatc acctgcagga caggcgcgcc ataacttcgt atagcataca
     4861 ttatacgaag ttatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag
     4921 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     4981 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     5041 gaccctttta gtcagtgtgg aaaatctcta gcacccgggc gattaaggaa agggctagat
     5101 cattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     5161 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5221 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5281 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5341 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5401 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5461 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5521 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     5581 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5641 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5701 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5761 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5821 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5881 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5941 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6001 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6061 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6121 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6181 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6241 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6301 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6361 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6421 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6481 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6541 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6601 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6661 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6721 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6781 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6841 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6901 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6961 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     7021 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     7081 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     7141 gcgcgttggc cgattcatta atgcagcaag ctcatggctg actaattttt tttatttatg
     7201 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     7261 gaggcctagg cttttgcaaa aagctccccg tggcacgaca ggtttcccga ctggaaagcg
     7321 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     7381 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     7441 ggaaacagct atgacatgat tacgaatttc acaaataaag catttttttc actgcattct
     7501 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcaa ctggataact
     7561 caagctaacc aaaatcatcc caaacttccc accccatacc ctattaccac tgccaattac
     7621 ctgtggtttc atttactcta aacctgtgat tcctctgaat tattttcatt ttaaagaaat
     7681 tgtatttgtt aaatatgtac tacaaactta gtagt
//
