LOCUS       Exported                8324 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Lentiviral vector, expresses EGFP under Oct3/4 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    PL-SIN-Oct4-EGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8324)
  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 8324)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8324
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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"
     regulatory      4759..4768
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4765..5484
                     /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             4765..5481
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4810..4831)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5071..5090)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5418..5439
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5491..5507
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5491..5507
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     LTR             5611..5844
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5881..5903)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5892..5908
                     /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(5916..5946)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5961..5982
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6099..6116)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6270..6858)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6350..6369)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7651..7670
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7889..7993)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        complement(8030..8226)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8044..8179
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8098..8117)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     8297..8315
                     /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 acggatcagc
     2761 tttgtgaact tggcggcttc caagtcgctg cctttattta ggtcttccaa ctaacctatg
     2821 gcactgttcc acaatgaatg tatagaaatt gggaggtgag catgacagag tggaggaaac
     2881 ggaagattca tggagagggc cagagagatg gcccctcagc caccctgggg gatgacttgg
     2941 acccatgtgg tagaaggagg ggacttccac acatgtgcta tgtgtagctg tgtgtaggta
     3001 catacacacc cttaaaataa aacgcaattt ttttttcaaa gtctcagggt gaatttggtg
     3061 aagtcgatga agctgaggca ggagaattat caggagttca agggcagctt gttttataga
     3121 gaaaggttcc atctctacct gatgaagact accatcaaga gacacccccg ccccccaggg
     3181 cacctagagc cactgaccct agccaacagc tcaggcgggc tgggcccagg ctcagaactc
     3241 tgtcctggct atgtacactg tggggtgctc tgggcttttt gaggctgtgt gattcaccct
     3301 ggggccttcg ttcagagcat ggtgtaggag cagacagaca aacaccatcc cttgcagaca
     3361 ggcactctga gggctattct cttgcaaaga taactaagca ccaggccagt aatgggatcc
     3421 tcagactggg cccagaaaac cactctaggg aagttcaggg taggctctct gcaccccctc
     3481 ctcctaatcc cgtctcctta gtgtctttcc gccagcacag gaatggggga ggggtgggtg
     3541 acgaggatga acaccggagt ccctggagga agggaagcag ggtatctcca tctgaggctc
     3601 tgtctttgag gagaggtgga gagctgggga agtcttgtgt gaggggattg gggctcagga
     3661 gggggttggg gagcaggaag ttgtccccag gggagccatc ctggcccatt caagggttga
     3721 gtacttgttt agggttagag ctgccccctc tggggaccag gattgtccag ccaaggccat
     3781 tgtcctgccc ccttccccca gtccctccca ggcccctttg aacctgaagt cagatatttc
     3841 ttctctctac ccacctccca cccgttgggt ttctccaccc aggaactagg ctggaagcct
     3901 gggatgagga ggtgggggga gggagaactg agaatcttga ggaaagaggc cccggcctta
     3961 actgtgaggg gatggagcct gggtgcaggt cttatggggg ttggggcggg tggttagtgt
     4021 ctaatctacc aacctggaca acacaagatg gaatactgtg ctctgaaaac gcagagccag
     4081 cacttctctg gggtctctgg ggacatatct ggttggggct cggggtccca tggtgtagag
     4141 cctctaaact ctggaggact ggaggtgcaa tggctgtctt gtcctggcct tggacatggg
     4201 ctgaaatact gggttcaccc atatctagga ctctagacgg gtgggtaagc aagaactgag
     4261 gagtggcccc agaaataatt ggcacacgaa cattcaatgg atgttttagg ctctccagag
     4321 gatggctgag tgggctgtaa ggacaggccg agagggtgca gtgccaacag gctttgtggt
     4381 gcgatggggc atccgagcaa ctggtttgtg aggtgtccgg tgacccaagg caggggtgag
     4441 aggaccttga aggttgaaaa tgaaggcctc ctggggtccc gtcctaaggg ttgtcctgtc
     4501 cagacgtccc caacctccgt ctggaagaca caggcagata gcgctcgcct cagtttctcc
     4561 cacccccaca gctctgctcc tccacccacc cagggggcgg ggccagaggt caaggctaga
     4621 gggtgggatt ggggagggag aggtgaaacc gtccctaggt gagccgtctt tccaccaggc
     4681 ccccggctcg gggtgcccac cttccccatg acgcgaattc tgcagtcgac ggtaccgcgg
     4741 gcccgggatc caccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
     4801 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     4861 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     4921 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     4981 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     5041 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     5101 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     5161 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     5221 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     5281 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     5341 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     5401 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     5461 ggcatggacg agctgtacaa gtaaagcgac tctagaacta gtggatcccc cgggctgcag
     5521 gaattcgata tcaagctggg gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     5581 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caaagaagac
     5641 aagatctgct ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc
     5701 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     5761 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     5821 agtcagtgtg gaaaatctct agcagaattc tagcttggcg taatcatggt catagctgtt
     5881 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5941 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     6001 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     6061 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     6121 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     6181 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6241 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6301 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6361 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6421 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     6481 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6541 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6601 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6661 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     6721 ggtatctgcg ctctgctgaa gccacttacc ttcggaaaaa gagttggtag ctcttgatcc
     6781 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6841 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6901 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6961 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     7021 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     7081 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     7141 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     7201 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     7261 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     7321 ttgcgcaacg ttgtgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7381 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7441 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7501 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7561 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7621 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7681 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7741 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7801 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7861 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7921 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7981 taggggttcc gcgcacattt ccccgaaaag tgccacctgg gactagcttt ttgcaaaagc
     8041 ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct
     8101 cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg
     8161 gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga
     8221 gatgtagctt gcatgccgac atggattatt gactagtccc taagaaacca ttattatcat
     8281 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtc
//
