LOCUS       sequence-9633-ex        7542 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 7542)
  AUTHORS   Soldner F, Hockemeyer D, Beard C, Gao Q, Bell GW, Cook EG, Hargus G,
            Blak A, Cooper O, Mitalipova M, Isacson O, Jaenisch R
  TITLE     Parkinson's disease patient-derived induced pluripotent stem cells 
            free of viral reprogramming factors.
  JOURNAL   Cell. 2009 Mar 6. 136(5):964-77.
  PUBMED    19269371
REFERENCE   2  (bases 1 to 7542)
  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: "Cell. 2009 
            Mar 6. 136(5):964-77."
COMMENT     Sequence Label: FUW-tetO-lox-hOCT4
FEATURES             Location/Qualifiers
     source          1..7542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        6..385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..585
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     540..560
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             603..783
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    830..955
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1448..1681
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1866..1910
                     /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    2208..2325
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    join(2384..2402,2403..2425,2426..2444,2445..2467,2468..2486,
                     2487..2509,2510..2528,2529..2551,2552..2570,2571..2593,
                     2594..2612,2613..2635,2636..2654)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2687..2724
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2721..2745
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     misc_feature    3931..4519
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3984..4004)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4402..4413)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4522..4538)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4523..4539)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    5043..5076
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             5090..5270
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5296..5313)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5302..5413
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(5559..6147)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5639..5658)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6318..7109,7110..7178))
                     /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     6941..6960
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7179..7283)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7358..7377)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
       61 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      121 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      181 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      361 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      421 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      481 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      541 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagcgcgt tttgcctgta
      601 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
      661 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
      721 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
      781 gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg
      841 caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac
      901 gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat
      961 taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa
     1021 aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa
     1081 tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc
     1141 ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg
     1201 tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga
     1261 gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag
     1321 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac
     1381 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag
     1441 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag
     1501 cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga
     1561 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca
     1621 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc
     1681 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta
     1741 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca
     1801 gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc
     1861 aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt
     1921 ttaacataac aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg
     1981 taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt
     2041 caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa
     2101 tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg aacggatcgg
     2161 cactgcgtgc gccaattctg cagacaaatg gcagtattca tccacaattt taaaagaaaa
     2221 ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata
     2281 caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg
     2341 gacagcagag atccagtttg gttaatgatc tcgagtttac cactccctat cagtgataga
     2401 gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt
     2461 ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt
     2521 gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag
     2581 tcgagtttac cactccctat cagtgataga gaaaagtgaa agtcgagttt accactccct
     2641 atcagtgata gagaaaagtg aaagtcgagc tcggtacccg ggtcgagtag gcgtgtacgg
     2701 tgggaggcct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
     2761 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg cggccccgaa
     2821 ttcatggcgg gacacctggc ttcggatttc gccttctcgc cccctccagg tggtggaggt
     2881 gatgggccag gggggccgga gccgggctgg gttgatcctc ggacctggct aagcttccaa
     2941 ggccctcctg gagggccagg aatcgggccg ggggttgggc caggctctga ggtgtggggg
     3001 attcccccat gccccccgcc gtatgagttc tgtgggggga tggcgtactg tgggccccag
     3061 gttggagtgg ggctagtgcc ccaaggcggc ttggagacct ctcagcctga gggcgaagca
     3121 ggagtcgggg tggagagcaa ctccgatggg gcctccccgg agccctgcac cgtcacccct
     3181 ggtgccgtga agctggagaa ggagaagctg gagcaaaacc cggaggagtc ccaggacatc
     3241 aaagctctgc agaaagaact cgagcaattt gccaagctcc tgaagcagaa gaggatcacc
     3301 ctgggatata cacaggccga tgtggggctc accctggggg ttctatttgg gaaggtattc
     3361 agccaaacga ccatctgccg ctttgaggct ctgcagctta gcttcaagaa catgtgtaag
     3421 ctgcggccct tgctgcagaa gtgggtggag gaagctgaca acaatgaaaa tcttcaggag
     3481 atatgcaaag cagaaaccct cgtgcaggcc cgaaagagaa agcgaaccag tatcgagaac
     3541 cgagtgagag gcaacctgga gaatttgttc ctgcagtgcc cgaaacccac actgcagcag
     3601 atcagccaca tcgcccagca gcttgggctc gagaaggatg tggtccgagt gtggttctgt
     3661 aaccggcgcc agaagggcaa gcgatcaagc agcgactatg cacaacgaga ggattttgag
     3721 gctgctgggt ctcctttctc agggggacca gtgtcctttc ctctggcccc agggccccat
     3781 tttggtaccc caggctatgg gagccctcac ttcactgcac tgtactcctc ggtccctttc
     3841 cctgaggggg aagcctttcc ccctgtctcc gtcaccactc tgggctctcc catgcattca
     3901 aactgagaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa
     3961 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     4021 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     4081 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     4141 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     4201 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     4261 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     4321 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     4381 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     4441 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     4501 ctttgggccg cctccccgca tcgataccgt cgacctcgag acctagaaaa acatggagca
     4561 atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag
     4621 gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag
     4681 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     4741 tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct
     4801 gattggcaga actacacacc agggccaggg atcagatatc cactgacctt tggatggtgc
     4861 tacaagctag taccagttga gcaagagaag gtagaagaag ccaatgaagg agagaacacc
     4921 cgcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga agtattagag
     4981 tggaggtttg acagccgcct agcatttcat cacatggccc gagagctgca tccggactcg
     5041 agataacttc gtataatgta tgctatacga agttattccg gactgtactg ggtctctctg
     5101 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     5161 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     5221 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gggcccgttt
     5281 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     5341 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     5401 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     5461 aggacagcaa gggggaggat tgggaagaca atagcaggca tgtgagcaaa aggccagcaa
     5521 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5581 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5641 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5701 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5761 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5821 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5881 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5941 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     6001 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6061 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6121 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6181 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6241 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6301 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6361 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6421 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6481 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6541 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6601 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6661 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6721 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6781 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6841 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6901 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6961 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7021 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7081 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7141 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7201 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7261 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     7321 tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat
     7381 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
     7441 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg
     7501 ttaggcgttt tgcgctgctt cgcgatgtac gggccagata ta
//
