LOCUS       sequence-9632-ex        7976 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 7976)
  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 7976)
  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-hKLF4
FEATURES             Location/Qualifiers
     source          1..7976
                     /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    4365..4953
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4418..4438)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4836..4847)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4956..4972)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4957..4973)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    5477..5510
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             5524..5704
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5730..5747)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5736..5847
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(5993..6581)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6073..6092)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6752..7543,7544..7612))
                     /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     7375..7394
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7613..7717)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7792..7811)
                     /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 attccttctc ttcgttgact ttggggttca ggtgccccag ctgcttcggg ctgccgagga
     2881 ccttctgggc ccccacatta atgaggcagc cacctggcga gtctgacatg gctgtcagcg
     2941 acgcgctgct cccatctttc tccacgttcg cgtctggccc ggcgggaagg gagaagacac
     3001 tgcgtcaagc aggtgccccg aataaccgct ggcgggagga gctctcccac atgaagcgac
     3061 ttcccccagt gcttcccggc cgcccctatg acctggcggc ggcgaccgtg gccacagacc
     3121 tggagagcgg cggagccggt gcggcttgcg gcggtagcaa cctggcgccc ctacctcgga
     3181 gagagaccga ggagttcaac gatctcctgg acctggactt tattctctcc aattcgctga
     3241 cccatcctcc ggagtcagtg gccgccaccg tgtcctcgtc agcgtcagcc tcctcttcgt
     3301 cgtcgccgtc gagcagcggc cctgccagcg cgccctccac ctgcagcttc acctatccga
     3361 tccgggccgg gaacgacccg ggcgtggcgc cgggcggcac gggcggaggc ctcctctatg
     3421 gcagggagtc cgctccccct ccgacggctc ccttcaacct ggcggacatc aacgacgtga
     3481 gcccctcggg cggcttcgtg gccgagctcc tgcggccaga attggacccg gtgtacattc
     3541 cgccgcagca gccgcagccg ccaggtggcg ggctgatggg caagttcgtg ctgaaggcgt
     3601 cgctgagcgc ccctggcagc gagtacggca gcccgtcggt catcagcgtc agcaaaggca
     3661 gccctgacgg cagccacccg gtggtggtgg cgccctacaa cggcgggccg ccgcgcacgt
     3721 gccccaagat caagcaggag gcggtctctt cgtgcaccca cttgggcgct ggaccccctc
     3781 tcagcaatgg ccaccggccg gctgcacacg acttccccct ggggcggcag ctccccagca
     3841 ggactacccc gaccctgggt cttgaggaag tgctgagcag cagggactgt caccctgccc
     3901 tgccgcttcc tcccggcttc catccccacc cggggcccaa ttacccatcc ttcctgcccg
     3961 atcagatgca gccgcaagtc ccgccgctcc attaccaaga gctcatgcca cccggttcct
     4021 gcatgccaga ggagcccaag ccaaagaggg gaagacgatc gtggccccgg aaaaggaccg
     4081 ccacccacac ttgtgattac gcgggctgcg gcaaaaccta cacaaagagt tcccatctca
     4141 aggcacacct gcgaacccac acaggtgaga aaccttacca ctgtgactgg gacggctgtg
     4201 gatggaaatt cgcccgctca gatgaactga ccaggcacta ccgtaaacac acggggcacc
     4261 gcccgttcca gtgccaaaaa tgcgaccgag cattttccag gtcggaccac ctcgccttac
     4321 acatgaagag gcatttttaa gaattcgata tcaagcttat cgataatcaa cctctggatt
     4381 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     4441 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     4501 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     4561 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     4621 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     4681 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     4741 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
     4801 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     4861 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     4921 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgacct cgagacctag
     4981 aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc
     5041 tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac
     5101 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     5161 aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac
     5221 aaggctactt ccctgattgg cagaactaca caccagggcc agggatcaga tatccactga
     5281 cctttggatg gtgctacaag ctagtaccag ttgagcaaga gaaggtagaa gaagccaatg
     5341 aaggagagaa cacccgcttg ttacaccctg tgagcctgca tgggatggat gacccggaga
     5401 gagaagtatt agagtggagg tttgacagcc gcctagcatt tcatcacatg gcccgagagc
     5461 tgcatccgga ctcgagataa cttcgtataa tgtatgctat acgaagttat tccggactgt
     5521 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     5581 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     5641 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     5701 agcagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     5761 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     5821 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     5881 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgtgag
     5941 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6001 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6061 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6121 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6181 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6241 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6301 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6361 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6421 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6481 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6541 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6601 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6661 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6721 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6781 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6841 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6901 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6961 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7021 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7081 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7141 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7201 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7261 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7321 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7381 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7441 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7501 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7561 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7621 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7681 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     7741 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
     7801 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
     7861 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
     7921 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatata
//
