LOCUS       sequence-121737-        4889 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EGFP under MERVL_LTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4889)
  AUTHORS   Ishiuchi T, Enriquez-Gasca R, Mizutani E, Boskovic A, 
            Ziegler-Birling C, Rodriguez-Terrones D, Wakayama T, Vaquerizas JM, 
            Torres-Padilla ME
  TITLE     Early embryonic-like cells are induced by downregulating 
            replication-dependent chromatin assembly.
  JOURNAL   Nat Struct Mol Biol. 2015 Aug 3. doi: 10.1038/nsmb.3066.
  PUBMED    26237512
REFERENCE   2  (bases 1 to 4889)
  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 Struct 
            Mol Biol. 2015 Aug 3. doi: 10.1038/nsmb.3066."
COMMENT     Sequence Label: 2C-EGFP
FEATURES             Location/Qualifiers
     source          1..4889
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      829..838
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(835..837,838..840,841..1554)
                     /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(835..837,838..840,841..1551)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(880..901)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1141..1160)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1488..1509
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1677..1798
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1714..1733)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1768..1787
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1805..2260)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1942..1963)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2154..2173
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2287..2391
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2393..2750
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2393..2584
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2416..2436)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2601..2736
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2785..3579
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2785..3579
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2839..2858)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3449..3468
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3767..3786)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3811..3858
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4187..4775
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4676..4695
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattat ggtcgacctg caggcggccg cgaattcact agtgattgcg gccgctgtag
       61 tggttattcc tggttgtcaa cttgacaata tttggaatga actacaattc ggaattggaa
      121 ggctcaccag tgacccttat ctggaggctt ggagatcctt atctggatct tggtttgaag
      181 atcttgagcc atagtggcta tggattccag aagattgaat ctccgagtta aggaacacac
      241 ctttaatctg ggctatgcct ttcatctggg attaaaggtg tggtggaaca cacctttaat
      301 ctgggctaca ccttttgctg gagacaatat aaggacattg gaagaaggga gtctagctct
      361 tgctcctgct ccttcgcctg cttgctgcgt gagactgagt aactactaga tccttggact
      421 tccattcaca gctgtgactg aacgattgtt gggaattgag ctgccgactg taagtcatca
      481 ataaattcct ttattatcta gagactattc ataagttctg tgactctaga gaaccctgac
      541 taatacagaa gttggtacca ggagtggttc tagagtaaca gaagtacaag gatgaatctt
      601 ttaaaatact ggaattggct tgttgatcca ccagcacttt caactattga aacctctcca
      661 gattctctcc ctcctgggag ctcagagaat tttgaagacc catggttgaa actctattct
      721 gaacttaaag aagctaatga ccttgatttt cttaatgaat taggtgattc agtgcacaaa
      781 gcttgtcgac aatcgaattc ccgcgggccc gggatccacc ggccggtcgc caccatggtg
      841 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
      901 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
      961 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     1021 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     1081 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     1141 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1201 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1261 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     1321 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     1381 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1441 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1501 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc
     1561 cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     1621 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     1681 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     1741 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag
     1801 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     1861 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     1921 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     1981 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     2041 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     2101 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     2161 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     2221 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     2281 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2341 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     2401 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2461 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     2521 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     2581 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     2641 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     2701 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     2761 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     2821 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     2881 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     2941 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     3001 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3061 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     3121 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3181 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     3241 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     3301 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     3361 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     3421 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     3481 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     3541 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     3601 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     3661 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     3721 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     3781 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     3841 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     3901 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     3961 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     4021 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4081 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4141 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4201 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4261 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4321 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4381 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4441 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4501 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4561 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4621 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4681 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4741 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4801 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     4861 ctgtggataa ccgtattacc gccatgcat
//
