LOCUS       sequence-350210-        7744 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7744)
  AUTHORS   Nishio H, Walsh MJ
  TITLE     CCAAT displacement protein/cut homolog recruits G9a histone lysine 
            methyltransferase to repress transcription.
  JOURNAL   Proc Natl Acad Sci U S A. 2004 Aug 3;101(31):11257-62. Epub 2004 Jul
            21.
  PUBMED    15269344
REFERENCE   2  (bases 1 to 7744)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2004-08-3"; volume: "101(31)"; pages: 
            "11257-62. Epub 2004 Jul 21"
COMMENT     Sequence Label: pEGFP-hG9a
FEATURES             Location/Qualifiers
     source          1..7744
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..305
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..509
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     460..480
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      548..557
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(554..556,557..559,560..1270)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(554..556,557..559,560..1270)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(599..620)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(860..879)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1207..1228
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4473..4594
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4510..4529)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4564..4583
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4601..5056)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4738..4759)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4950..4969
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5083..5187
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5189..5546
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5189..5380
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5212..5232)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5397..5532
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5581..6375
                     /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             5581..6375
                     /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(5635..5654)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6245..6264
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6563..6582)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6607..6654
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6983..7571
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7472..7491
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
       61 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      121 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      181 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      241 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      301 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      361 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      421 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      481 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct
      541 accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
      601 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
      661 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
      721 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
      781 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
      841 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
      901 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
      961 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     1021 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     1081 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     1141 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     1201 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     1261 gctgtacaag tccggactca gatctcgagc tcaagcttcg aattctatga gtgatgatgt
     1321 ccactcactg ggaaaggtga cctcagatct ggccaaaagg aggaagctga actcaggagg
     1381 tggcctgtcg gaggagttag gttctgcccg gcgttcagga gaagtgaccc tgacgaaagg
     1441 ggaccccggg tccctggagg agtgggagac ggtggtgggt gatgacttca gtctctacta
     1501 tgattcctac tctgtggatg agcgcgtgga ctccgacagc aagtctgaag ttgaagctct
     1561 aactgaacaa ctaagtgaag aggaggagga ggaagaggag gaagaagaag aagaggaaga
     1621 ggaggaggaa gaggaagaag aagaggaaga tgaggagtca gggaatcagt cagataggag
     1681 tggttccagt ggccggcgca aggccaagaa gaaatggcga aaagacagcc catgggtgaa
     1741 gccgtctcgg aaacggcgca agcgggagcc tccgcgggcc aaggagccac gaggagtgaa
     1801 tggtgtgggc tcctcaggcc ccagtgagta catggaggtc cctctggggt ccctggagct
     1861 gcccagcgag gggaccctct cccccaacca cgctggggtg tccaatgaca catcttcgct
     1921 ggagacagag cgagggtttg aggagttgcc cctgtgcagc tgccgcatgg aggcacccaa
     1981 gattgaccgc atcagcgaga gggcggggca caagtgcatg gccactgaga gtgtggacgg
     2041 agagctgtca ggctgcaatg ccgccatcct caagcgggag accatgaggc catccagccg
     2101 tgtggccctg atggtgctct gtgagaccca ccgcgcccgc atggtcaaac accactgctg
     2161 cccgggctgc ggctacttct gcacggcggg caccttcctg gagtgccacc ctgacttccg
     2221 tgtggcccac cgcttccaca aggcctgtgt gtctcagctg aatgggatgg tcttctgtcc
     2281 ccactgtggg gaggatgctt ctgaagctca agaggtgacc atcccccggg gtgacggggt
     2341 gaccccaccg gccggcactg cagctcctgc acccccaccc ctgtcccagg atgtccccgg
     2401 gagagcagac acttctcagc ccagtgcccg gatgcgaggg catggggaac cccggcgccc
     2461 gccctgcgat cccctggctg acaccattga cagctcaggg ccctccctga ccctgcccaa
     2521 tgggggctgc ctttcagccg tggggctgcc actggggcca ggccgggagg ccctggaaaa
     2581 ggccctggtc atccaggagt cagagaggcg gaagaagctc cgtttccacc ctcggcagtt
     2641 gtacctgtcc gtgaagcagg gcgagctgca gaaggtgatc ctgatgctgt tggacaacct
     2701 ggaccccaac ttccagagcg accagcagag caagcgcacg cccctgcatg cagccgccca
     2761 gaagggctcc gtggagatct gccatgtgct gctgcaggct ggagccaaca taaatgcagt
     2821 ggacaaacag cagcggacgc cactgatgga ggccgtggtg aacaaccacc tggaggtagc
     2881 ccgttacatg gtgcagcgtg gtggctgtgt ctatagcaag gaggaggacg gttccacctg
     2941 cctccaccac gcagccaaaa tcgggaactt ggagatggtc agcctgctgc tgagcacagg
     3001 acaggtggac gtcaacgccc aggacagtgg ggggtggacg cccatcatct gggctgcaga
     3061 gcacaagcac atcgaggtga tccgcatgct actgacgcgg ggcgccgacg tcaccctcac
     3121 tgacaacgag gagaacatct gcctgcactg ggcctccttc acgggcagcg ccgccatcgc
     3181 cgaagtcctt ctgaatgcgc gctgtgacct ccatgctgtc aactaccatg gggacacccc
     3241 cctgcacatc gcagctcggg agagctacca tgactgcgtg ctgttattcc tgtcacgtgg
     3301 ggccaaccct gagctgcgga acaaagaggg ggacacagca tgggacctga ctcccgagcg
     3361 ctccgacgtg tggtttgcgc ttcaactcaa ccgcaagctc cgacttgggg tgggaaatcg
     3421 ggccatccgc acagagaaga tcatctgccg ggacgtggct cggggctatg agaacgtgcc
     3481 cattccctgt gtcaacggtg tggatgggga gccctgccct gaggattaca agtacatctc
     3541 agagaactgc gagacgtcca ccatgaacat cgatcgcaac atcacccacc tgcagcactg
     3601 cacgtgtgtg gacgactgct ctagctccaa ctgcctgtgc ggccagctca gcatccggtg
     3661 ctggtatgac aaggatgggc gattgctcca ggaatttaac aagattgagc ctccgctgat
     3721 tttcgagtgt aaccaggcgt gctcatgctg gagaaactgc aagaaccggg tcgtacagag
     3781 tggcatcaag gtgcggctac agctctaccg aacagccaag atgggctggg gggtccgcgc
     3841 cctgcagacc atcccacagg ggaccttcat ctgcgagtat gtcggggagc tgatctctga
     3901 tgctgaggct gatgtgagag aggatgattc ttacctcttc gacttagaca acaaggatgg
     3961 agaggtgtac tgcatagatg cccgttacta tggcaacatc agccgcttca tcaaccacct
     4021 gtgtgacccc aacatcattc ccgtccgggt cttcatgctg caccaagacc tgcgatttcc
     4081 acgcatcgcc ttcttcagtt cccgagacat ccggactggg gaggagctag ggtttgacta
     4141 tggcgaccgc ttctgggaca tcaaaagcaa atatttcacc tgccaatgtg gctctgagaa
     4201 gtgcaagcac tcagccgaag ccattgccct ggagcagagc cgtctggccc gcctggaccc
     4261 acaccctgag ctgctgcccg agctcggctc cctgccccct gtcaacacat gagaattctg
     4321 cagtcgacgg taccgcgggc ccgggatcca ccggatctag ataactgatc ataatcagcc
     4381 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     4441 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     4501 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4561 gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt
     4621 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     4681 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     4741 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     4801 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     4861 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     4921 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     4981 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     5041 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     5101 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5161 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     5221 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5281 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     5341 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     5401 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5461 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     5521 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     5581 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     5641 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     5701 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     5761 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     5821 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     5881 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     5941 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     6001 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     6061 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     6121 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     6181 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     6241 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     6301 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     6361 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     6421 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     6481 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     6541 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     6601 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     6661 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     6721 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     6781 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     6841 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6901 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6961 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     7021 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     7081 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     7141 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     7201 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     7261 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     7321 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     7381 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     7441 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     7501 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     7561 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     7621 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
     7681 tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     7741 ttcc
//
