LOCUS       sequence-9566-ex        9852 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 9852)
  AUTHORS   Carey BW, Markoulaki S, Hanna J, Saha K, Gao Q, Mitalipova M, 
            Jaenisch R
  TITLE     Reprogramming of murine and human somatic cells using a single 
            polycistronic vector.
  JOURNAL   Proc Natl Acad Sci U S A. 2009 Jan 6. 106(1):157-62.
  PUBMED    19109433
REFERENCE   2  (bases 1 to 9852)
  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: "Proc Natl 
            Acad Sci U S A. 2009 Jan 6. 106(1):157-62."
COMMENT     Sequence Label: FUW-KO
FEATURES             Location/Qualifiers
     source          1..9852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1686..1919
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2104..2148
                     /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    2446..2563
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     2622..2638
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2680..2713)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2763..3974
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3811..3830
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5479..5544
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     protein_bind    complement(6652..6685)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    6741..7329
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6794..6814)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7212..7223)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7332..7348)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             7858..8038
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(8100..8688)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8180..8199)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8859..9650,9651..9719))
                     /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     9482..9501
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9720..9824)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg gccgcgctga tcttcagacc tggaggagga
     1561 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
     1621 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
     1681 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
     1741 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
     1801 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1861 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1921 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1981 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     2041 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     2101 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     2161 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     2221 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     2281 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     2341 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatcggca
     2401 ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg
     2461 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     2521 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     2581 cagcagagat ccagtttggt tagtaccggg cccgctctag tcgaggtcga cggtatcgat
     2641 aagctcgctt cacgagattc cagcaggtcg agggacctaa taacttcgta tagcatacat
     2701 tatacgaagt tatattaagg gttccaagct taagcggcct aacccgtgtc ggctccagat
     2761 ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcctca cggcgagcgc
     2821 tgccacgtca gacgaagggc gcagcgagcg tcctgatcct tccgcccgga cgctcaggac
     2881 agcggcccgc tgctcataag actcggcctt agaaccccag tatcagcaga aggacatttt
     2941 aggacgggac ttgggtgact ctagggcact ggttttcttt ccagagagcg gaacaggcga
     3001 ggaaaagtag tcccttctcg gcgattctgc ggagggatct ccgtggggcg gtgaacgccg
     3061 atgattatat aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg
     3121 gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg
     3181 ctggccgggg ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg tggagagacc
     3241 gccaagggct gtagtctggg tccgcgagca aggttgccct gaactggggg ttggggggag
     3301 cgcagcaaaa tggcggctgt tcccgagtct tgaatggaag acgcttgtga ggcgggctgt
     3361 gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg tcttgaggcc
     3421 ttcgctaatg cgggaaagct cttattcggg tgagatgggc tggggcacca tctggggacc
     3481 ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt cgtctgttgc gggggcggca
     3541 gttatggcgg tgccgttggg cagtgcaccc gtacctttgg gagcgcgcgc cctcgtcgtg
     3601 tcgtgacgtc acccgttctg ttggcttata atgcagggtg gggccacctg ccggtaggtg
     3661 tgcggtaggc ttttctccgt cgcaggacgc agggttcggg cctagggtag gctctcctga
     3721 atcgacaggc gccggacctc tggtgagggg agggataagt gaggcgtcag tttctttggt
     3781 cggttttatg tacctatctt cttaagtagc tgaagctccg gttttgaact atgcgctcgg
     3841 ggttggcgag tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa tcatttgggt
     3901 caatatgtaa ttttcagtgt tagactagta aattgtccgc taaattctgg ccgtttttgg
     3961 cttttttgtt agacgaagct tgggctgcag gtcgactcta gagaattcgc catgaggcag
     4021 ccacctggcg agtctgacat ggctgtcagc gacgctctgc tcccgtcctt ctccacgttc
     4081 gcgtccggcc cggcgggaag ggagaagaca ctgcgtccag caggtgcccc gactaaccgt
     4141 tggcgtgagg aactctctca catgaagcga cttcccccac ttcccggccg cccctacgac
     4201 ctggcggcga cggtggccac agacctggag agtggcggag ctggtgcagc ttgcagcagt
     4261 aacaacccgg ccctcctagc ccggagggag accgaggagt tcaacgacct cctggaccta
     4321 gactttatcc tttccaactc gctaacccac caggaatcgg tggccgccac cgtgaccacc
     4381 tcggcgtcag cttcatcctc gtcttcccca gcgagcagcg gccctgccag cgcgccctcc
     4441 acctgcagct tcagctatcc gatccgggcc gggggtgacc cgggcgtggc tgccagcaac
     4501 acaggtggag ggctcctcta cagccgagaa tctgcgccac ctcccacggc ccccttcaac
     4561 ctggcggaca tcaatgacgt gagcccctcg ggcggcttcg tggctgagct cctgcggccg
     4621 gagttggacc cagtatacat tccgccacag cagcctcagc cgccaggtgg cgggctgatg
     4681 ggcaagtttg tgctgaaggc gtctctgacc acccctggca gcgagtacag cagcccttcg
     4741 gtcatcagtg ttagcaaagg aagcccagac ggcagccacc ccgtggtagt ggcgccctac
     4801 agcggtggcc cgccgcgcat gtgccccaag attaagcaag aggcggtccc gtcctgcacg
     4861 gtcagccggt ccctagaggc ccatttgagc gctggacccc agctcagcaa cggccaccgg
     4921 cccaacacac acgacttccc cctggggcgg cagctcccca ccaggactac ccctacactg
     4981 agtcccgagg aactgctgaa cagcagggac tgtcaccctg gcctgcctct tcccccagga
     5041 ttccatcccc atccggggcc caactaccct cctttcctgc cagaccagat gcagtcacaa
     5101 gtcccctctc tccattatca agagctcatg ccaccgggtt cctgcctgcc agaggagccc
     5161 aagccaaaga ggggaagaag gtcgtggccc cggaaaagaa cagccaccca cacttgtgac
     5221 tatgcaggct gtggcaaaac ctataccaag agttctcatc tcaaggcaca cctgcgaact
     5281 cacacaggcg agaaacctta ccactgtgac tgggacggct gtgggtggaa attcgcccgc
     5341 tccgatgaac tgaccaggca ctaccgcaaa cacacagggc accggccctt tcagtgccag
     5401 aagtgcgaca gggccttttc caggtcggac caccttgcct tacacatgaa gaggcacttt
     5461 gctagcggca gcggcccggt gaaacagact ttgaattttg accttctcaa gttggcggga
     5521 gacgtcgagt ccaaccctgg gcccgcatgc accatggctg gacacctggc ttcagacttc
     5581 gccttctcac ccccaccagg tgggggtgat gggtcagcag ggctggagcc gggctgggtg
     5641 gatcctcgaa cctggctaag cttccaaggg cctccaggtg ggcctggaat cggaccaggc
     5701 tcagaggtat tggggatctc cccatgtccg cccgcatacg agttctgcgg agggatggca
     5761 tactgtggac ctcaggttgg actgggccta gtcccccaag ttggcgtgga gactttgcag
     5821 cctgagggcc aggcaggagc acgagtggaa agcaactcag agggaacctc ctctgagccc
     5881 tgtgccgacc gccccaatgc cgtgaagttg gagaaggtgg aaccaactcc cgaggagtcc
     5941 caggacatga aagccctgca gaaggagcta gaacagtttg ccaagctgct gaagcagaag
     6001 aggatcacct tggggtacac ccaggccgac gtggggctca ccctgggcgt tctctttgga
     6061 aaggtgttca gccagaccac catctgtcgc ttcgaggcct tgcagctcag ccttaagaac
     6121 atgtgtaagc tgcggcccct gctggagaag tgggtggagg aagccgacaa caatgagaac
     6181 cttcaggaga tatgcaaatc ggagaccctg gtgcaggccc ggaagagaaa gcgaactagc
     6241 attgagaacc gtgtgaggtg gagtctggag accatgtttc tgaagtgccc gaagccctcc
     6301 ctacagcaga tcactcacat cgccaatcag cttgggctag agaaggatgt ggttcgagta
     6361 tggttctgta accggcgcca gaagggcaaa agatcaagta ttgagtattc ccaacgagaa
     6421 gagtatgagg ctacagggac acctttccca gggggggctg tatcctttcc tctgccccca
     6481 ggtccccact ttggcacccc aggctatgga agcccccact tcaccacact ctactcagtc
     6541 ccttttcctg agggcgaggc ctttccctct gttcccgtca ctgctctggg ctctcccatg
     6601 cattcaaact gagcatgcct cgagggcgcg ccgaattcgt cgagggacct aataacttcg
     6661 tatagcatac attatacgaa gttatacatg tttaagggtt ccggttccac taggtacaat
     6721 tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg
     6781 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     6841 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     6901 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     6961 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     7021 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     7081 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     7141 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     7201 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     7261 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     7321 cctccccgca tcgataccgt cgacctcgat cgagacctag aaaaacatgg agcaatcaca
     7381 agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca agaggaggag
     7441 gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct
     7501 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     7561 cgaagacaag atatccttga tctgtggatc taccacacac aaggctactt ccctgattgg
     7621 cagaactaca caccagggcc agggatcaga tatccactga cctttggatg gtgctacaag
     7681 ctagtaccag ttgagcaaga gaaggtagaa gaagccaatg aaggagagaa cacccgcttg
     7741 ttacaccctg tgagcctgca tgggatggat gacccggaga gagaagtatt agagtggagg
     7801 tttgacagcc gcctagcatt tcatcacatg gcccgagagc tgcatccgga ctgtactggg
     7861 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     7921 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     7981 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagc
     8041 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     8101 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     8161 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     8221 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     8281 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     8341 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     8401 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     8461 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     8521 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     8581 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     8641 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     8701 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     8761 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     8821 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     8881 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     8941 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     9001 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     9061 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     9121 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     9181 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     9241 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     9301 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     9361 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     9421 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     9481 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     9541 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     9601 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     9661 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     9721 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     9781 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     9841 gtgccacctg ac
//
