LOCUS       sequence-9862-ex        7813 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector, expresses EGFP under Nanog promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7813)
  AUTHORS   Hotta A, Cheung AY, Farra N, Vijayaragavan K, Seguin CA, Draper JS, 
            Pasceri P, Maksakova IA, Mager DL, Rossant J, Bhatia M, Ellis J
  TITLE     Isolation of human iPS cells using EOS lentiviral vectors to select 
            for pluripotency.
  JOURNAL   Nat Methods. 2009 May . 6(5):370-6.
  PUBMED    19404254
REFERENCE   2  (bases 1 to 7813)
  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: "Nat 
            Methods. 2009 May . 6(5):370-6."
COMMENT     Sequence Label: PL-SIN-Nanog-EGFP
FEATURES             Location/Qualifiers
     source          1..7813
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             785..1042
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1693..1810
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    2200..2433
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             join(4254..4256,4257..4259,4260..4973)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4254..4256,4257..4259,4260..4970)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4299..4320)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4560..4579)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4907..4928
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             5100..5333
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5370..5392)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5381..5397
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5405..5411,5412..5429,5430..5435))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5450..5471
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5588..5605)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5759..6347)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5839..5858)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6816..7193)
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     primer_bind     7140..7159
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7378..7482)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        complement(7519..7715)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7533..7668
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7786..7804
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gcggccgctg gaagggctaa
      421 tttggtccca aaaaagacaa gagatccttg atctgtggat ctaccacaca caaggctact
      481 tccctgattg gcagaactac acaccagggc cagggatcag atatccactg acctttggat
      541 ggtgcttcaa gttagtacca gttgaaccag agcaagtaga agaggccaaa taaggagaga
      601 agaacagctt gttacaccct atgagccagc atgggatgga ggacccggag ggagaagtat
      661 tagtgtggaa gtttgacagc ctcctagcat ttcgtcacat ggcccgagag ctgcatccgg
      721 agtactacaa agactgctga catcgagctt tctacaaggg actttccgct ggggactttc
      781 cagggaggtg tggcctgggc gggactgggg agtggcgagc cctcagatgc tacatataag
      841 cagctgcttt ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
      901 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgctcaaa
      961 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     1021 tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagtaaagc
     1081 cagaggagat ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg
     1141 ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg
     1201 ggtgcgagag cgtcggtatt aagcggggga gaattagata aatgggaaaa aattcggtta
     1261 aggccagggg gaaagaaaca atataaacta aaacatatag tatgggcaag cagggagcta
     1321 gaacgattcg cagttaatcc tggcctttta gagacatcag aaggctgtag acaaatactg
     1381 ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca
     1441 atagcagtcc tctattgtgt gcatcaaagg atagatgtaa aagacaccaa ggaagcctta
     1501 gataagatag aggaagagca aaacaaaagt aagaaaaagg cacagcaagc agcagctgac
     1561 acaggaaaca acagccaggt cagccaaaat taccctatag tgcagaacct ccaggggcaa
     1621 atggtacatc aggccatatc acctagaact ttaaattaag acagcagtac aaatggcagt
     1681 attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat
     1741 agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat
     1801 tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggaaag gaccagcaaa
     1861 gctcctctgg aaaggtgaag gggcagtagt aatacaagat aatagtgaca taaaagtagt
     1921 gccaagaaga aaagcaaaga tcatcaggga ttatggaaaa cagatggcag gtgatgattg
     1981 tgtggcaagt agacaggatg aggattaaca catggaaaag attagtaaaa caccatagct
     2041 ctagagcgat ccgatcttca gacctggagg aggcgatatg agggacaatt ggagaagtga
     2101 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
     2161 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
     2221 cttgggagca gcaggaagca ctatgggctg cacgtcaatg acgctgacgg tacaggccag
     2281 acaattattg tctgatatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
     2341 acagcatctg ttgcaactca cagtctgggg catcaaacag ctccaggcaa gaatcctggc
     2401 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
     2461 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
     2521 ttggaataac atgacctgga tggagtggga cagagaaatt aacaattaca caagcttggt
     2581 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
     2641 accattatcg tttcagaccc acctcccaat cccgagggga cccgacaggc ccgaaggaat
     2701 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatccca
     2761 aaagtcagct tgtgtgggag caaagccagc tgctctgggt gcagacccag gagcagagtg
     2821 cagaggagaa tgagtcaaag agttttgtct tcaaaaatta cataatcggg atttgctaag
     2881 agtttacttt tcggtatgga agactggaaa agagaaagaa atcttaggtt tcttgaatgt
     2941 tgggtttggg aataggaagg aaaatcgaaa actgtagact ttgtccataa atgttagtgc
     3001 tggaacccca ctctaaaaac tttgttcctt tggaaaacac ctcccttccc ccagaaacac
     3061 acacacccac acgagatggg cacggagtag tcttgaaaga catgacaaat caccagacct
     3121 gggaagaagc taaagagcca gagggaaaaa gccagaagtc gactacctgg gaggagggat
     3181 agacaagaaa ccaaactaaa ggaaactaag gtaggtgctg aaaacaagta ccattttcaa
     3241 cattaactga tgccttggct tcatgctata atgccatgtt gtgtttcact ataacctcag
     3301 agtgaatgaa agaggaaaat ggagctagtt gaaatttctg cctaaactag ccagattttg
     3361 agacactaag ttatctcaaa tcaagaaatc accctaatga gaatttcaat aacctcagga
     3421 atttaaggtg catgcatccc ccaccccccc cttttttttt tgagacgtag tcccgctctg
     3481 ttgcccaggc tggagtacag tggcgcgata tcggctcacc acaacctctg cctcccaggt
     3541 tcaagggatt ctcccgcctc agcttccaga gtagctggga ctacagacac ccaccaccat
     3601 gcgtggctaa tttttgtatt tttagtagag agggggtttc gccatgttgg ccaggctggt
     3661 ttcaaactcc tgacttcagg tgatccgcct gccacggcct cccaatttac tgggattaca
     3721 ggggtgggcc accgcgcccg gcctttttct taatttttaa aaatattaaa gttttatccc
     3781 attcctgttg aaccatattc ctgatttaaa agttggaaac gtggtgaacc tagaagtatt
     3841 tgttgctggg tttgtcttca ggttctgttg ctcggttttc tagttcccca cctagtctgg
     3901 gttactctgc agctactttt gcattacaat ggccttggtg agactggtag acgggattaa
     3961 ctgagaattc acaagggtgg gtcagtaggg ggtgtgcccg ccaggagggg tgggtctaag
     4021 gtgatagagc cttcattata aatctagaga ctccaggatt ttaacgttct gctggactga
     4081 gctggttgcc tcatgttatt atgcaggcaa ctcactttat cccaatttct tgatactttt
     4141 ccttctggag gtcctatttc tctaacatct tccagaaaag tcttaaagct gccttaacct
     4201 tttttccagt ccacctctta aattttttcc tcctcttcct ctatactaac accatggtga
     4261 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     4321 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     4381 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     4441 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     4501 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     4561 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     4621 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     4681 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     4741 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     4801 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     4861 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     4921 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taaagcggcc
     4981 aacctcgacg ttattccctt cgcgaaggcc tatcgatttc gaacccgggg tacctttaag
     5041 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact
     5101 ggaagggcta attcactccc aaagaagaca agatctgctt tttgcctgta ctgggtctct
     5161 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     5221 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     5281 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagaattct
     5341 agcttggcgt aatcatggtc atagctgttc ctgtgtgaaa ttgttatccg ctcacaattc
     5401 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5461 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5521 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5581 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5641 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5701 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5761 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5821 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5881 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5941 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6001 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6061 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6121 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6181 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccacttacct
     6241 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6301 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6361 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6421 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6481 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6541 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6601 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6661 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6721 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6781 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgtgccattg ctacaggcat
     6841 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6901 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6961 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7021 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7081 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7141 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7201 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7261 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7321 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7381 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7441 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7501 gccacctggg actagctttt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt
     7561 ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa aaattagtca
     7621 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
     7681 gggcgggact atggttgctg actaattgag atgtagcttg catgccgaca tggattattg
     7741 actagtccct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     7801 aggccctttc gtc
//
