LOCUS       sequence-483086-        8343 bp    DNA     circular SYN 13-AUG-2025
DEFINITION  Lentiviral vector, expresses EGFP under Oct3/4 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8343)
  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 8343)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 13, 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-Oct4-EGFP
FEATURES             Location/Qualifiers
     source          1..8343
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             78..258
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    909..1026
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    1417..1650
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     regulatory      3974..3983
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3980..3982,3983..3985,3986..4699)
                     /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(3980..3982,3983..3985,3986..4696)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4025..4046)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4286..4305)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4633..4654
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4716..4732
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4716..4732
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     LTR             4836..5069
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5103..5119)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5103..5119)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5116..5138)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5127..5143
                     /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(5151..5157,5158..5175,5176..5181))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5196..5217
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5334..5351)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5505..6093)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5585..5604)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6264..7055,7056..7124))
                     /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     6887..6906
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7125..7229)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7280..7415
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7532..7550
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7588..7610)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7710..7729
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7923..7945
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7937..7954
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7938..7954
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gaggtgtggc ctgggcggga ctggggagtg gcgagccctc agatgctaca tataagcagc
       61 tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
      121 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg ctcaaagtag
      181 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
      241 tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag taaagccaga
      301 ggagatctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg
      361 gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg
      421 cgagagcgtc ggtattaagc gggggagaat tagataaatg ggaaaaaatt cggttaaggc
      481 cagggggaaa gaaacaatat aaactaaaac atatagtatg ggcaagcagg gagctagaac
      541 gattcgcagt taatcctggc cttttagaga catcagaagg ctgtagacaa atactgggac
      601 agctacaacc atcccttcag acaggatcag aagaacttag atcattatat aatacaatag
      661 cagtcctcta ttgtgtgcat caaaggatag atgtaaaaga caccaaggaa gccttagata
      721 agatagagga agagcaaaac aaaagtaaga aaaaggcaca gcaagcagca gctgacacag
      781 gaaacaacag ccaggtcagc caaaattacc ctatagtgca gaacctccag gggcaaatgg
      841 tacatcaggc catatcacct agaactttaa attaagacag cagtacaaat ggcagtattc
      901 atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
      961 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
     1021 aattttcggg tttattacag ggacagcaga gatccagttt ggaaaggacc agcaaagctc
     1081 ctctggaaag gtgaaggggc agtagtaata caagataata gtgacataaa agtagtgcca
     1141 agaagaaaag caaagatcat cagggattat ggaaaacaga tggcaggtga tgattgtgtg
     1201 gcaagtagac aggatgagga ttaacacatg gaaaagatta gtaaaacacc atagctctag
     1261 agcgatcccg atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt
     1321 atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag
     1381 aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt
     1441 gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca
     1501 attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca
     1561 gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt
     1621 ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat
     1681 ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg
     1741 gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa gcttggtagg
     1801 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     1861 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     1921 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcagcttt
     1981 gtgaacttgg cggcttccaa gtcgctgcct ttatttaggt cttccaacta acctatggca
     2041 ctgttccaca atgaatgtat agaaattggg aggtgagcat gacagagtgg aggaaacgga
     2101 agattcatgg agagggccag agagatggcc cctcagccac cctgggggat gacttggacc
     2161 catgtggtag aaggagggga cttccacaca tgtgctatgt gtagctgtgt gtaggtacat
     2221 acacaccctt aaaataaaac gcaatttttt tttcaaagtc tcagggtgaa tttggtgaag
     2281 tcgatgaagc tgaggcagga gaattatcag gagttcaagg gcagcttgtt ttatagagaa
     2341 aggttccatc tctacctgat gaagactacc atcaagagac acccccgccc cccagggcac
     2401 ctagagccac tgaccctagc caacagctca ggcgggctgg gcccaggctc agaactctgt
     2461 cctggctatg tacactgtgg ggtgctctgg gctttttgag gctgtgtgat tcaccctggg
     2521 gccttcgttc agagcatggt gtaggagcag acagacaaac accatccctt gcagacaggc
     2581 actctgaggg ctattctctt gcaaagataa ctaagcacca ggccagtaat gggatcctca
     2641 gactgggccc agaaaaccac tctagggaag ttcagggtag gctctctgca ccccctcctc
     2701 ctaatcccgt ctccttagtg tctttccgcc agcacaggaa tgggggaggg gtgggtgacg
     2761 aggatgaaca ccggagtccc tggaggaagg gaagcagggt atctccatct gaggctctgt
     2821 ctttgaggag aggtggagag ctggggaagt cttgtgtgag gggattgggg ctcaggaggg
     2881 ggttggggag caggaagttg tccccagggg agccatcctg gcccattcaa gggttgagta
     2941 cttgtttagg gttagagctg ccccctctgg ggaccaggat tgtccagcca aggccattgt
     3001 cctgccccct tcccccagtc cctcccaggc ccctttgaac ctgaagtcag atatttcttc
     3061 tctctaccca cctcccaccc gttgggtttc tccacccagg aactaggctg gaagcctggg
     3121 atgaggaggt ggggggaggg agaactgaga atcttgagga aagaggcccc ggccttaact
     3181 gtgaggggat ggagcctggg tgcaggtctt atgggggttg gggggtggtt agtgtctaat
     3241 ctaccaacct ggacaacaca agatggaata ctgtgctctg aaaacgcaga gccagcactt
     3301 ctctggggtc tctggggaca tatctggttg gggctcgggg tcccatggtg tagagcctct
     3361 aaactctgga ggactggagg tgcaatggct gtcttgtcct ggccttggac atgggctgaa
     3421 atactgggtt cacccatatc taggactcta gacgggtggg taagcaagaa ctgaggagtg
     3481 gccccagaaa taattggcac acgaacattc aatggatgtt ttaggctctc cagaggatgg
     3541 ctgagtgggc tgtaaggaca ggccgagagg gtgcagtgcc aacaggcttt gtggtgcgat
     3601 ggggcatccg agcaactggt ttgtgaggtg tccggtgacc caaggcaggg gtgagaggac
     3661 cttgaaggtt gaaaatgaag gcctcctggg gtcccgtcct aagggttgtc ctgtccagac
     3721 gtccccaacc tccgtctgga agacacaggc agatagcgct cgcctcagtt tctcccaccc
     3781 ccacagctct gctcctccac ccacccaggg ggcggggcca gaggtcaagg ctagagggtg
     3841 ggattgggga gggagaggtg aaaccgtccc taggtgagcc gtctttccac caggcccccg
     3901 gctcggggtg cccaccttcc ccatgacgcg aattctgcag tcgacggtac cgcgggcccg
     3961 ggatccaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     4021 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     4081 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     4141 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     4201 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     4261 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     4321 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     4381 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     4441 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     4501 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     4561 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     4621 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     4681 ggacgagctg tacaagtaaa gcggccggcc gcgactctag aactagtgga tcccccgggc
     4741 tgcaggaatt cgatatcaag ctggggtacc tttaagacca atgacttaca aggcagctgt
     4801 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaaag
     4861 aagacaagat ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg
     4921 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     4981 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     5041 cttttagtca gtgtggaaaa tctctagcag aattctagcg attctagctt ggcgtaatca
     5101 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5161 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5221 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5281 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5341 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5401 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5461 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5521 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5581 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5641 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5701 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5761 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5821 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5881 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5941 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6001 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6061 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6121 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6181 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6241 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6301 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6361 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6421 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6481 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6541 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6601 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6661 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6721 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6781 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6841 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6901 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6961 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7021 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7081 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7141 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7201 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgggact
     7261 agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca
     7321 gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga
     7381 gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg
     7441 gttgctgact aattgagatg agcttgcatg ccgacattga ttattgacta gtccctaaga
     7501 aaccattctt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     7561 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     7621 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     7681 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     7741 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     7801 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     7861 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     7921 ttcccagtca cgacgttgta aaacgacggc cagtgaattg cggccgctgg aagggctaat
     7981 ttggtcccaa aaaagacaag agatccttga tctgtggatc taccacacac aaggctactt
     8041 ccctgattgg cagaactaca caccagggcc agggatcaga tatccactga cctttggatg
     8101 gtgcttcaag ttagtaccag ttgaaccaga gcaagtagaa gaggccaatg aaggagagaa
     8161 caacagcttg ttacacccta tgagccagca tgggatggag gacccggagg gagaagtatt
     8221 agtgtggaag tttgacagcc tcctagcatt tcgtcacatg gcccgagagc tgcatccgga
     8281 gtactacaaa gactgctgac atcgagcttt ctacaaggga ctttccgctg gggactttcc
     8341 agg
//
