LOCUS       sequence-9634-ex        7420 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 7420)
  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 7420)
  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-hSOX2
FEATURES             Location/Qualifiers
     source          1..7420
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        7..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        388..586
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     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
                     /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    2209..2326
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    join(2385..2403,2404..2426,2427..2445,2446..2468,2469..2487,
                     2488..2510,2511..2529,2530..2552,2553..2571,2572..2594,
                     2595..2613,2614..2636,2637..2655)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /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"
     misc_feature    3809..4397
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3862..3882)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4280..4291)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4400..4416)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4401..4417)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    4921..4954
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             4968..5148
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5174..5191)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5180..5291
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(5437..6025)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5517..5536)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6196..6987,6988..7056))
                     /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     6819..6838
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7057..7161)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7236..7255)
                     /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 attcgccacc atgtacaaca tgatggagac ggagctgaag ccgccgggcc cgcagcaaac
     2881 ttcggggggc ggcggcggca actccaccgc ggcggcggcc ggcggcaacc agaaaaacag
     2941 cccggaccgc gtcaagcggc ccatgaatgc cttcatggtg tggtcccgcg ggcagcggcg
     3001 caagatggcc caggagaacc ccaagatgca caactcggag atcagcaagc gcctgggcgc
     3061 cgagtggaaa cttttgtcgg agacggagaa gcggccgttc atcgacgagg ctaagcggct
     3121 gcgagcgctg cacatgaagg agcacccgga ttataaatac cggccccggc ggaaaaccaa
     3181 gacgctcatg aagaaggata agtacacgct gcccggcggg ctgctggccc ccggcggcaa
     3241 tagcatggcg agcggggtcg gggtgggcgc cggcctgggc gcgggcgtga accagcgcat
     3301 ggacagttac gcgcacatga acggctggag caacggcagc tacagcatga tgcaggacca
     3361 gctgggctac ccgcagcacc cgggcctcaa tgcgcacggc gcagcgcaga tgcagcccat
     3421 gcaccgctac gacgtgagcg ccctgcagta caactccatg accagctcgc agacctacat
     3481 gaacggctcg cccacctaca gcatgtccta ctcgcagcag ggcacccctg gcatggctct
     3541 tggctccatg ggttcggtgg tcaagtccga ggccagctcc agcccccctg tggttacctc
     3601 ttcctcccac tccagggcgc cctgccaggc cggggacctc cgggacatga tcagcatgta
     3661 tctccccggc gccgaggtgc cggaacccgc cgcccccagc agacttcaca tgtcccagca
     3721 ctaccagagc ggcccggtgc ccggcacggc cattaacggc acactgcccc tctcacacat
     3781 gtgagaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag
     3841 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     3901 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     3961 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     4021 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     4081 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     4141 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     4201 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     4261 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     4321 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     4381 ttgggccgcc tccccgcatc gataccgtcg acctcgagac ctagaaaaac atggagcaat
     4441 cacaagtagc aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga
     4501 ggaggaggtg ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc
     4561 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     4621 ccaacgaaga caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga
     4681 ttggcagaac tacacaccag ggccagggat cagatatcca ctgacctttg gatggtgcta
     4741 caagctagta ccagttgagc aagagaaggt agaagaagcc aatgaaggag agaacacccg
     4801 cttgttacac cctgtgagcc tgcatgggat ggatgacccg gagagagaag tattagagtg
     4861 gaggtttgac agccgcctag catttcatca catggcccga gagctgcatc cggactcgag
     4921 ataacttcgt ataatgtatg ctatacgaag ttattccgga ctgtactggg tctctctggt
     4981 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     5041 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     5101 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagg gcccgtttaa
     5161 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     5221 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     5281 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     5341 gacagcaagg gggaggattg ggaagacaat agcaggcatg tgagcaaaag gccagcaaaa
     5401 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5461 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5521 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5581 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5641 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5701 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5761 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5821 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5881 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5941 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6001 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6061 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6121 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6181 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6241 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6301 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6361 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6421 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6481 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6541 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6601 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6661 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6721 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6781 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6841 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6901 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6961 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7021 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7081 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7141 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     7201 gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag
     7261 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     7321 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
     7381 aggcgttttg cgctgcttcg cgatgtacgg gccagatata
//
