LOCUS       sequence-121736-        6535 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 3XturboGFP-NLS-PEST under MERVL_LTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6535)
  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 6535)
  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-3XtbGFP-PEST
FEATURES             Location/Qualifiers
     source          1..6535
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             823..1518
                     /codon_start=1
                     /product="improved green fluorescent protein from 
                     Pontellina plumata (Evdokimov et al., 2006)"
                     /label=TurboGFP
                     /note="mammalian codon-optimized"
                     /translation="MESDESGLPAMEIECRITGTLNGVEFELVGGGEGTPEQGRMTNKM
                     KSTKGALTFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVL
                     HVSFSYRYEAGRVIGDFKVMGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNDLDGSFTR
                     TFSLRDGGYYSSVVDSHMHFKSAIHPSILQNGGPMFAFRRVEEDHSNTELGIVEYQHAF
                     KTPDADAGEE"
     CDS             1546..2241
                     /codon_start=1
                     /product="improved green fluorescent protein from 
                     Pontellina plumata (Evdokimov et al., 2006)"
                     /label=TurboGFP
                     /note="mammalian codon-optimized"
                     /translation="MESDESGLPAMEIECRITGTLNGVEFELVGGGEGTPEQGRMTNKM
                     KSTKGALTFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVL
                     HVSFSYRYEAGRVIGDFKVMGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNDLDGSFTR
                     TFSLRDGGYYSSVVDSHMHFKSAIHPSILQNGGPMFAFRRVEEDHSNTELGIVEYQHAF
                     KTPDADAGEE"
     CDS             2269..2964
                     /codon_start=1
                     /product="improved green fluorescent protein from 
                     Pontellina plumata (Evdokimov et al., 2006)"
                     /label=TurboGFP
                     /note="mammalian codon-optimized"
                     /translation="MESDESGLPAMEIECRITGTLNGVEFELVGGGEGTPEQGRMTNKM
                     KSTKGALTFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVL
                     HVSFSYRYEAGRVIGDFKVMGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNDLDGSFTR
                     TFSLRDGGYYSSVVDSHMHFKSAIHPSILQNGGPMFAFRRVEEDHSNTELGIVEYQHAF
                     KTPDADAGEE"
     CDS             2983..3003
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3007..3027
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3031..3051
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3323..3444
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3360..3379)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3414..3433
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3451..3906)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3588..3609)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3800..3819
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3933..4037
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4039..4396
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4039..4230
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4062..4082)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4247..4382
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4431..5225
                     /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             4431..5225
                     /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(4485..4504)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5095..5114
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5413..5432)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5457..5504
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5833..6421
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6322..6341
                     /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 ccgcggggag aaccctggac ctatggagag cgacgagagc
      841 ggcctgcccg ccatggagat cgagtgccgc atcaccggca ccctgaacgg cgtggagttc
      901 gagctggtgg gcggcggaga gggcaccccc gagcagggcc gcatgaccaa caagatgaag
      961 agcaccaaag gcgccctgac cttcagcccc tacctgctga gccacgtgat gggctacggc
     1021 ttctaccact tcggcaccta ccccagcggc tacgagaacc ccttcctgca cgccatcaac
     1081 aacggcggct acaccaacac ccgcatcgag aagtacgagg acggcggcgt gctgcacgtg
     1141 agcttcagct accgctacga ggccggccgc gtgatcggcg acttcaaggt gatgggcacc
     1201 ggcttccccg aggacagcgt gatcttcacc gacaagatca tccgcagcaa cgccaccgtg
     1261 gagcacctgc accccatggg cgataacgat ctggatggca gcttcacccg caccttcagc
     1321 ctgcgcgacg gcggctacta cagctccgtg gtggacagcc acatgcactt caagagcgcc
     1381 atccacccca gcatcctgca gaacgggggc cccatgttcg ccttccgccg cgtggaggag
     1441 gatcacagca acaccgagct gggcatcgtg gagtaccagc acgccttcaa gaccccggat
     1501 gcagatgccg gtgaagaact cagatccact ggaggcggag gtggaatgga gagcgacgag
     1561 agcggcctgc ccgccatgga gatcgagtgc cgcatcaccg gcaccctgaa cggcgtggag
     1621 ttcgagctgg tgggcggcgg agagggcacc cccgagcagg gccgcatgac caacaagatg
     1681 aagagcacca aaggcgccct gaccttcagc ccctacctgc tgagccacgt gatgggctac
     1741 ggcttctacc acttcggcac ctaccccagc ggctacgaga accccttcct gcacgccatc
     1801 aacaacggcg gctacaccaa cacccgcatc gagaagtacg aggacggcgg cgtgctgcac
     1861 gtgagcttca gctaccgcta cgaggccggc cgcgtgatcg gcgacttcaa ggtgatgggc
     1921 accggcttcc ccgaggacag cgtgatcttc accgacaaga tcatccgcag caacgccacc
     1981 gtggagcacc tgcaccccat gggcgataac gatctggatg gcagcttcac ccgcaccttc
     2041 agcctgcgcg acggcggcta ctacagctcc gtggtggaca gccacatgca cttcaagagc
     2101 gccatccacc ccagcatcct gcagaacggg ggccccatgt tcgccttccg ccgcgtggag
     2161 gaggatcaca gcaacaccga gctgggcatc gtggagtacc agcacgcctt caagaccccg
     2221 gatgcagatg ccggtgaaga actcagatcc actggaggcg gaggtggaat ggagagcgac
     2281 gagagcggcc tgcccgccat ggagatcgag tgccgcatca ccggcaccct gaacggcgtg
     2341 gagttcgagc tggtgggcgg cggagagggc acccccgagc agggccgcat gaccaacaag
     2401 atgaagagca ccaaaggcgc cctgaccttc agcccctacc tgctgagcca cgtgatgggc
     2461 tacggcttct accacttcgg cacctacccc agcggctacg agaacccctt cctgcacgcc
     2521 atcaacaacg gcggctacac caacacccgc atcgagaagt acgaggacgg cggcgtgctg
     2581 cacgtgagct tcagctaccg ctacgaggcc ggccgcgtga tcggcgactt caaggtgatg
     2641 ggcaccggct tccccgagga cagcgtgatc ttcaccgaca agatcatccg cagcaacgcc
     2701 accgtggagc acctgcaccc catgggcgat aacgatctgg atggcagctt cacccgcacc
     2761 ttcagcctgc gcgacggcgg ctactacagc tccgtggtgg acagccacat gcacttcaag
     2821 agcgccatcc accccagcat cctgcagaac gggggcccca tgttcgcctt ccgccgcgtg
     2881 gaggaggatc acagcaacac cgagctgggc atcgtggagt accagcacgc cttcaagacc
     2941 ccggatgcag atgccggtga agaactcaga tctcgagctg atccaaaaaa gaagagaaag
     3001 gtagatccaa aaaagaagag aaaggtagat ccaaaaaaga agagaaaggt aggatccacc
     3061 ggatctagcc atggcttccc gccggaggtg gaggagcagg atgatggcac gctgcccatg
     3121 tcttgtgccc aggagagcgg gatggaccgt caccctgcag cctgtgcttc tgctaggatc
     3181 aatgtgtagt gtacaagtaa agcggccgcg actctagatc ataatcagcc ataccacatt
     3241 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     3301 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     3361 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3421 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
     3481 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     3541 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     3601 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     3661 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     3721 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     3781 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     3841 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     3901 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3961 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4021 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     4081 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4141 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4201 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4261 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4321 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4381 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     4441 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4501 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4561 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     4621 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4681 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4741 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4801 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4861 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4921 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     4981 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     5041 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     5101 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     5161 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     5221 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     5281 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     5341 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     5401 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     5461 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     5521 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     5581 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     5641 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     5701 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5761 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5821 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5881 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5941 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     6001 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6061 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6121 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6181 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6241 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6301 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6361 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6421 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6481 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
