LOCUS       Exported                6446 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    WZLneo-Cre
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6446)
  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 6446)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6446
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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"
     CDS             1387..1407
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1408..2436
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     misc_feature    2486..2947
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2542..2559)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2769..2788
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2952..3746
                     /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(3006..3025)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3616..3635
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             3933..4402
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4532..4549)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4703..5291)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4783..4802)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5451..6311)
                     /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     6074..6093
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6312..6416)
                     /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 tgatatcatc gatgaattcg
     1381 accatgccca agaagaagag gaaggtgtcc aatttactga ccgtacacca aaatttgcct
     1441 gcattaccgg tcgatgcaac gagtgatgag gttcgcaaga acctgatgga catgttcagg
     1501 gatcgccagg cgttttctga gcatacctgg aaaatgcttc tgtccgtttg ccggtcgtgg
     1561 gcggcatggt gcaagttgaa taaccggaaa tggtttcccg cagaacctga agatgttcgc
     1621 gattatcttc tatatcttca ggcgcgcggt ctggcagtaa aaactatcca gcaacatttg
     1681 ggccagctaa acatgcttca tcgtcggtcc gggctgccac gaccaagtga cagcaatgct
     1741 gtttcactgg ttatgcggcg gatccgaaaa gaaaacgttg atgccggtga acgtgcaaaa
     1801 caggctctag cgttcgaacg cactgatttc gaccaggttc gttcactcat ggaaaatagc
     1861 gatcgctgcc aggatatacg taatctggca tttctgggga ttgcttataa caccctgtta
     1921 cgtatagccg aaattgccag gatcagggtt aaagatatct cacgtactga cggtgggaga
     1981 atgttaatcc atattggcag aacgaaaacg ctggttagca ccgcaggtgt agagaaggca
     2041 cttagcctgg gggtaactaa actggtcgag cgatggattt ccgtctctgg tgtagctgat
     2101 gatccgaata actacctgtt ttgccgggtc agaaaaaatg gtgttgccgc gccatctgcc
     2161 accagccagc tatcaactcg cgccctggaa gggatttttg aagcaactca tcgattgatt
     2221 tacggcgcta aggatgactc tggtcagaga tacctggcct ggtctggaca cagtgcccgt
     2281 gtcggagccg cgcgagatat ggcccgcgct ggagtttcaa taccggagat catgcaagct
     2341 ggtggctgga ccaatgtaaa tattgtcatg aactatatcc gtaacctgga tagtgaaaca
     2401 ggggcaatgg tgcgcctgct ggaagatggc gattagtcga ccaattccgg ttattttcca
     2461 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     2521 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     2581 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     2641 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     2701 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     2761 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     2821 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
     2881 ggttaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg
     2941 atgataatac catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3001 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3061 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3121 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3181 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     3241 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     3301 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     3361 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     3421 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     3481 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     3541 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     3601 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     3661 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     3721 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcatcga taaaataaaa
     3781 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa
     3841 gctagcttaa gtaacgccat tttgcaaggc atggaaaaat acataactga gaatagagaa
     3901 gttcagatca aggtcaggaa cagatggaac agctgaatat gggccaaaca ggatatctgt
     3961 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggaacagctg aatatgggcc
     4021 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc
     4081 ccagatgcgg tccagccctc agcagtttct agagaaccat cagatgtttc cagggtgccc
     4141 caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt
     4201 ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcgggg
     4261 cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc
     4321 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
     4381 acccgtcagc gggggtcttt cacatgcagc atgtatcaaa attaatttgg ttttttttct
     4441 taagtattta cattaaatgg ccatagttgc attaatgaat cggccaacgc gcggggagag
     4501 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4561 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4621 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4681 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4741 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4801 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4861 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4921 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4981 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5041 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5101 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5161 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5221 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5281 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5341 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5401 tgcggccgca aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5461 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5521 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5581 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5641 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5701 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5761 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5821 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5881 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5941 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6001 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6061 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6121 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6181 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6241 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6301 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6361 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6421 tttccccgaa aagtgccacc tgcagc
//
