LOCUS       sequence-202166-        8055 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8055)
  AUTHORS   Blelloch R, Venere M, Yen J, Ramalho-Santos M
  TITLE     Generation of induced pluripotent stem cells in the absence of drug 
            selection.
  JOURNAL   Cell Stem Cell. 2007 Sep 13;1(3):245-7. doi: 
            10.1016/j.stem.2007.08.008. Epub 2007 Sep 6.
  PUBMED    18371358
REFERENCE   2  (bases 1 to 8055)
  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.stem.2007.08.008"; journalName: "Cell Stem Cell"; date: 
            "2007-09-13- 13"; volume: "1"; issue: "3"; pages: "245-7"
COMMENT     Sequence Label: pLOVE-Klf4
FEATURES             Location/Qualifiers
     source          1..8055
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1447..1680
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1865..1909
                     /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    2207..2324
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2390..2693
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2694..2897
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    2899..2917
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2920..2938
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2979..3003
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4462..4486)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4539..4580
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    complement(4618..4651)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4707..5295
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4760..4780)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5178..5189)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5298..5314)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5299..5315)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5824..6004
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6066..6654)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6146..6165)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6825..7616,7617..7685))
                     /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     7448..7467
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7686..7790)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7865..7884)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg atcttcagac ctggaggagg
     1321 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc
     1381 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt
     1441 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc
     1501 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa
     1561 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat
     1621 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct
     1681 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
     1741 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag
     1801 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
     1861 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
     1921 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
     1981 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
     2041 accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat
     2101 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc
     2161 actgcgtgcg ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag
     2221 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2281 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2341 acagcagaga tccagtttgg ttagtaccgg gcccgctcta gcagttccgc gttacataac
     2401 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     2461 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     2521 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     2581 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     2641 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     2701 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     2761 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     2821 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     2881 tctatataag cagagctctc cctatcagtg atagagatct ccctatcagt gatagagatc
     2941 gtcgactagt ccagtgtggt ggaattctgc agatatcaac aagtttgtac aaaaaagcag
     3001 gctccgcggc cgcccccttc accatggctg tcagcgacgc tctgctcccg tccttctcca
     3061 cgttcgcgtc cggcccggcg ggaagggaga agacactgcg tccagcaggt gccccgacta
     3121 accgttggcg tgaggaactc tctcacatga agcgacttcc cccacttccc ggccgcccct
     3181 acgacctggc ggcgacggtg gccacagacc tggagagtgg cggagctggt gcagcttgca
     3241 gcagtaacaa cccggccctc ctagcccgga gggagaccga ggagttcaac gacctcctgg
     3301 acctagactt tatcctttcc aactcgctaa cccaccagga atcggtggcc gccaccgtga
     3361 ccacctcggc gtcagcttca tcctcgtctt ccccagcgag cagcggccct gccagcgcgc
     3421 cctccacctg cagcttcagc tatccgatcc gggccggggg tgacccgggc gtggctgcca
     3481 gcaacacagg tggagggctc ctctacagcc gagaatctgc gccacctccc acggccccct
     3541 tcaacctggc ggacatcaat gacgtgagcc cctcgggcgg cttcgtggct gagctcctgc
     3601 ggccggagtt ggacccagta tacattccgc cacagcagcc tcagccgcca ggtggcgggc
     3661 tgatgggcaa gtttgtgctg aaggcgtctc tgaccacccc tggcagcgag tacagcagcc
     3721 cttcggtcat cagtgttagc aaaggaagcc cagacggcag ccaccccgtg gtagtggcgc
     3781 cctacagcgg tggcccgccg cgcatgtgcc ccaagattaa gcaagaggcg gtcccgtcct
     3841 gcacggtcag ccggtcccta gaggcccatt tgagcgctgg accccagctc agcaacggcc
     3901 accggcccaa cacacacgac ttccccctgg ggcggcagct ccccaccagg actaccccta
     3961 cactgagtcc cgaggaactg ctgaacagca gggactgtca ccctggcctg cctcttcccc
     4021 caggattcca tccccatccg gggcccaact accctccttt cctgccagac cagatgcagt
     4081 cacaagtccc ctctctccat tatcaagagc tcatgccacc gggttcctgc ctgccagagg
     4141 agcccaagcc aaagagggga agaaggtcgt ggccccggaa aagaacagcc acccacactt
     4201 gtgactatgc aggctgtggc aaaacctata ccaagagttc tcatctcaag gcacacctgc
     4261 gaactcacac aggcgagaaa ccttaccact gtgactggga cggctgtggg tggaaattcg
     4321 cccgctcgga tgaactgacc aggcactacc gcaaacacac agggcaccgg ccctttcagt
     4381 gccagaagtg cgacagggcc ttttccaggt cggaccacct tgccttacac atgaagaggc
     4441 actttaaggg tgggcgcgcc gacccagctt tcttgtacaa agtggttgat atccagcaca
     4501 gtggcggccg ctcgagtcta gagggcccgc ggttcgaagg taagcctatc cctaaccctc
     4561 tcctcggtct cgattctacg cgtaccggtt agtaatgaca attcgtcgag ggacctaata
     4621 acttcgtata gcatacatta tacgaagtta tacatgttta agggttccgg ttccactagg
     4681 tacaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
     4741 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     4801 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     4861 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     4921 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     4981 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     5041 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     5101 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     5161 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     5221 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     5281 gggccgcctc cccgcatcga taccgtcgac ctcgatcgag acctagaaaa acatggagca
     5341 atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag
     5401 gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag
     5461 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     5521 tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct
     5581 gattggcaga actacacacc agggccaggg atcagatatc cactgacctt tggatggtgc
     5641 tacaagctag taccagttga gcaagagaag gtagaagaag ccaatgaagg agagaacacc
     5701 cgcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga agtattagag
     5761 tggaggtttg acagccgcct agcatttcat cacatggccc gagagctgca tccggactgt
     5821 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     5881 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     5941 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     6001 agcagcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     6061 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     6121 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     6181 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     6241 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     6301 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     6361 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     6421 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6481 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     6541 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6601 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6661 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6721 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6781 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6841 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6901 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6961 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     7021 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     7081 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     7141 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     7201 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     7261 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     7321 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     7381 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     7441 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     7501 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     7561 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7621 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7681 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7741 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7801 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc
     7861 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
     7921 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
     7981 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg
     8041 ccagatatac gcgtt
//
