LOCUS       sequence-197039-        6528 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 6528)
  AUTHORS   Wang L, Jin Q, Lee JE, Su IH, Ge K
  TITLE     Histone H3K27 methyltransferase Ezh2 represses Wnt genes to 
            facilitate adipogenesis.
  JOURNAL   Proc Natl Acad Sci U S A. 2010 Apr 20. 107(16):7317-22.
  PUBMED    20368440
REFERENCE   2  (bases 1 to 6528)
  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. 2010 Apr 20. 107(16):7317-22."
COMMENT     Sequence Label: WZLneo-C/EBP-alpha
FEATURES             Location/Qualifiers
     source          1..6528
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..470
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    535..734
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             935..1351
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1281..1303
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1319..1335
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1382..1391
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2568..3029
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2624..2641)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2851..2870
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3034..3828
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3088..3107)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3698..3717
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             4015..4484
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4614..4631)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4785..5373)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4865..4884)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5533..6324,6325..6393))
                     /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     6156..6175
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6394..6498)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
       61 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
      121 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
      181 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
      241 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
      301 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
      361 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
      421 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct
      481 cgtccgggat cgggagaccc ctgcccaggg accaccgacc caccaccggg aggcaagctg
      541 gccagcaact tatctgtgtc tgtccgattg tctagtgtct atgactgatt ttatgcgcct
      601 gcgtcggtac tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga
      661 gttctgaaca cccggccgca accctgggag acgtcccagg gactttgggg gccgtttttg
      721 tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat atgtggttct
      781 ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg
      841 gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt gttgtctctg
      901 tctgactgtg tttctgtatt tgtctgaaaa ttagggccag actgttacca ctcccttaag
      961 tttgacctta ggtcactgga aagatgtcga gcggatcgct cacaaccagt cggtagatgt
     1021 caagaagaga cgttgggtta ccttctgctc tgcagaatgg ccaaccttta acgtcggatg
     1081 gccgcgagac ggcaccttta accgagacct catcacccag gttaagatca aggtcttttc
     1141 acctggcccg catggacacc cagaccaggt cccctacatc gtgacctggg aagccttggc
     1201 ttttgacccc cctccctggg tcaagccctt tgtacaccct aagcctccgc ctcctcttcc
     1261 tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct
     1321 ttatccagcc ctcactcctt ctctaggcgc cggccggatc ccccgggctg caggaattcg
     1381 cgccaccatg gagtcggccg acttctacga ggcggagccg cggcccccga tgagcagcca
     1441 cctccagagc cccccgcacg cgcccagcaa cgccgccttt ggctttcccc ggggcgcggg
     1501 ccccgcgccg cccccagccc cacctgccgc cccggagccg ctgggcggca tctgcgagca
     1561 cgagacgtct atagacatca gcgcctacat cgacccggcc gccttcaacg acgagttcct
     1621 ggccgacctc ttccagcaca gccggcagca ggagaaggcc aaggcggcgg cgggccccgc
     1681 gggtggcggc ggtgactttg actacccggg cgccccggcg ggccccggcg gtgcggtcat
     1741 gtccgcgggg gcgcacggac cccctcccgg ctacggctgt gcggcggccg gctacctgga
     1801 cggcaggctg gagcccctgt acgagcgcgt cggggcgccc gcgctgcggc cgctggtgat
     1861 caagcaggag ccccgcgagg aggacgaggc gaagcagctg gcgctggccg gcctcttccc
     1921 ctatcagccc ccgccgccgc cgccgccacc gcacccgcac gcgtctcccg cgcacttggc
     1981 cgcccctcac ttgcagttcc agatcgcaca ctgcggccag accaccatgc acctgcagcc
     2041 tggccaccct acgccgccgc cgacgcccgt gcccagccct catcccgcgc ctgcaatggg
     2101 tgctgcgggc ctgccgggcc ccgggggctc gctcaagggc ttggctggtc cgcaccccga
     2161 cctccgcacc ggcggcggcg gcggcggcgg ggccggcgcg ggcaaggcca agaagtcggt
     2221 ggataagaac agcaacgagt accgggtacg gcgggaacgc aacaacatcg cggtgcgcaa
     2281 gagccgagat aaagccaaac agcgcaacgt ggagacgcag cagaaggtgt tggagttgac
     2341 cagtgacaat gaccgcctgc gcaagcgggt ggaacagctg agccgtgaac tggacacgct
     2401 gcggggtatc ttccgccagc tgcctgagag ctccttggtc aaggccatgg gcaactgcgc
     2461 gtgaggcgcg aattcgccag cacagtggtc gagatacgta aaacaattga agatctagtc
     2521 gaccaattcc ggttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
     2581 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
     2641 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca
     2701 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
     2761 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
     2821 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
     2881 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca
     2941 tgctttacat gtgtttagtc gaggttaaaa aacgtctagg ccccccgaac cacggggacg
     3001 tggttttcct ttgaaaaaca cgatgataat accatgattg aacaagatgg attgcacgca
     3061 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     3121 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     3181 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     3241 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     3301 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     3361 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     3421 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     3481 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     3541 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     3601 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     3661 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     3721 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     3781 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     3841 ggttcgcatc gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag
     3901 accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa
     3961 atacataact gagaatagag aagttcagat caaggtcagg aacagatgga acagctgaat
     4021 atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag
     4081 atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     4141 cagggccaag aacagatggt ccccagatgc ggtccagccc tcagcagttt ctagagaacc
     4201 atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa
     4261 ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga
     4321 gcccacaacc cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg
     4381 tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag
     4441 ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcacatgca gcatgtatca
     4501 aaattaattt ggtttttttt cttaagtatt tacattaaat ggccatagtt gcattaatga
     4561 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4621 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4681 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4741 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4801 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4861 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4921 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4981 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5041 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5101 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5161 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5221 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5281 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5341 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5401 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5461 aggatcttca cctagatcct tttgcggccg caaatcaatc taaagtatat atgagtaaac
     5521 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5581 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5641 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5701 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5761 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5821 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5881 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5941 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6001 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6061 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6121 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6181 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6241 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6301 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6361 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6421 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6481 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgcagc
//
