LOCUS       Exported                8019 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pMSCVhygro-Cre
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8019)
  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 8019)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8019
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    55..76
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        91..121
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    129..145
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     LTR             307..821
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     CDS             complement(1179..2204)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(2222..2721)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2270..2289)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     2663..2686
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     CDS             complement(2743..3771)
                     /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"
     CDS             complement(3772..3792)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(3830..4246)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(3846..3862)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(3878..3900)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    4313..4654
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             4718..5234
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(5434..5453)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(5581..5603)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5797..5816)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5916..5938
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5976..5994)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6062..6166
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6167..7027
                     /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     complement(6385..6404)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7198..7786
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7687..7706
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7940..7957
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
       61 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      121 gtgtggaatt gtgagcggat aacaatttca caattctccc cagctctttc tgcgctgtcc
      181 ataatgaact atttcaggag tattggagta tgtggattca aaacagtggc taattgagtc
      241 aggattactt aatattcgac tcctaccctc agaatgttgt tctccaactt caagaaactg
      301 ttacccatga aagacccccg ctgacgggta gtcaatcact cagaggagac cctcccaagg
      361 aacagcgaga ccacaagtcg gatgcaactg caagagggtt tattggatac acgggtaccc
      421 gggcgacgca gtctatcgga ggactggcgc gccgagtgag gggttgtggg ctcttttatt
      481 gagctcgggg agcagaagcg cgcgaacaga agcgagaagc gaactgattg gttagttcaa
      541 ataaggcaca gggtcatttc aggtccttgg ggcaccctgg aaacatctga tggttctcta
      601 gaaactgctg agggcgggac cgcatctggg gaccatctgt tcttggccct gagccggggc
      661 aggaactgct taccacagat atcctgtttg gcccatattc tgctgtctct ctgttcctaa
      721 ccttgatctg aacttctcta ttctcagtta tgtattttcc atgccttgca aaatggcgtt
      781 acttaagcta gcttgccaaa cctacaggtg gggtctttca ttcccccctt tttctggaga
      841 ctaaataaaa tcttttattt tatcgataag cttggctgca ggtcgacgga tctgcaccca
      901 atcggcaggc acgggcggcg atctccaatc tgcgggatca gtcagatcac ccgagtgcgt
      961 gggcatgaca atcgtgccct ggggaccaac acaatccaga agggcctgaa tcactgcgac
     1021 cggccctccc gcgacccagc cgagcgagct tagcgaactg tggacgagaa ctgtgccacc
     1081 aagcgtaagg ccgttctctc gcatttgcct tgctaggctc gcgcgagttg ctggctgagg
     1141 cgttctcgaa atcagctctt gttcggtcgg catctactct attcctttgc cctcggacga
     1201 gtgctggggc gtcggtttcc actatcggcg agtacttcta cacagccatc ggtccagacg
     1261 gccgcgcttc tgcgggcgat ttgtgtacgc ccgacagtcc cggctccgga tcggacgatt
     1321 gcgtcgcatc gaccctgcgc ccaagctgca tcatcgaaat tgccgtcaac caagctctga
     1381 tagagttggt caagaccaat gcggagcata tacgcccgga gccgcggcga tcctgcaagc
     1441 tccggatgcc tccgctcgaa gtagcgcgtc tgctgctcca tacaagccaa ccacggcctc
     1501 cagaagaaga tgttggcgac ctcgtattgg gaatccccga acatcgcctc gctccagtca
     1561 atgaccgctg ttatgcggcc attgtccgtc aggacattgt tggagccgaa atccgcgtgc
     1621 acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca tcagctcatc gagagcctgc
     1681 gcgacggacg cactgacggt gtcgtccatc acagtttgcc agtgatacac atggggatca
     1741 gcaatcgcgc atatgaaatc acgccatgta gtgtattgac cgattccttg cggtccgaat
     1801 gggccgaacc cgctcgtctg gctaagatcg gccgcagcga tcgcatccat ggcctccgcg
     1861 accggctgca gaacagcggg cagttcggtt tcaggcaggt cttgcaacgt gacaccctgt
     1921 gcacggcggg agatgcaata ggtcaggctc tcgctgaatt ccccaatgtc aagcacttcc
     1981 ggaatcggga gcgcggccga tgcaaagtgc cgataaacat aacgatcttt gtagaaacca
     2041 tcggcgcagc tatttacccg caggacatat ccacgccctc ctacatcgaa gctgaaagca
     2101 cgagattctt cgccctccga gagctgcatc aggtcggaga cgctgtcgaa cttttcgatc
     2161 agaaacttct cgacagacgt cgcggtgagt tcaggctttt tcatggtggc gggatgcagg
     2221 tcgaaaggcc cggagatgag gaagaggaga acagcgcggc agacgtgcgc ttttgaagcg
     2281 tgcagaatgc cgggcctccg gaggaccttc gggcgcccgc cccgcccctg agcccgcccc
     2341 tgagcccgcc cccggaccca ccccttccca gcctctgagc ccagaaagcg aaggagcaaa
     2401 gctgctattg gccgctgccc caaaggccta cccgcttcca ttgctcagcg gtgctgtcca
     2461 tctgcacgag actagtgaga cgtgctactt ccatttgtca cgtcctgcac gacgcgagct
     2521 gcggggcggg ggggaacttc ctgactaggg gaggagtaga aggtggcgcg aaggggccac
     2581 caaagaacgg agccggttgg cgcctaccgg tggatgtgga atgtgtgcga ggccagaggc
     2641 cacttgtgta gcgccaagtg cccagcgggg ctgctaaagc gcatgctcca gactgccttg
     2701 ggaaaagcgc ctcccctacc cggtagaatt cgttaacctc gactaatcgc catcttccag
     2761 caggcgcacc attgcccctg tttcactatc caggttacgg atatagttca tgacaatatt
     2821 tacattggtc cagccaccag cttgcatgat ctccggtatt gaaactccag cgcgggccat
     2881 atctcgcgcg gctccgacac gggcactgtg tccagaccag gccaggtatc tctgaccaga
     2941 gtcatcctta gcgccgtaaa tcaatcgatg agttgcttca aaaatccctt ccagggcgcg
     3001 agttgatagc tggctggtgg cagatggcgc ggcaacacca ttttttctga cccggcaaaa
     3061 caggtagtta ttcggatcat cagctacacc agagacggaa atccatcgct cgaccagttt
     3121 agttaccccc aggctaagtg ccttctctac acctgcggtg ctaaccagcg ttttcgttct
     3181 gccaatatgg attaacattc tcccaccgtc agtacgtgag atatctttaa ccctgatcct
     3241 ggcaatttcg gctatacgta acagggtgtt ataagcaatc cccagaaatg ccagattacg
     3301 tatatcctgg cagcgatcgc tattttccat gagtgaacga acctggtcga aatcagtgcg
     3361 ttcgaacgct agagcctgtt ttgcacgttc accggcatca acgttttctt ttcggatccg
     3421 ccgcataacc agtgaaacag cattgctgtc acttggtcgt ggcagcccgg accgacgatg
     3481 aagcatgttt agctggccca aatgttgctg gatagttttt actgccagac cgcgcgcctg
     3541 aagatataga agataatcgc gaacatcttc aggttctgcg ggaaaccatt tccggttatt
     3601 caacttgcac catgccgccc acgaccggca aacggacaga agcattttcc aggtatgctc
     3661 agaaaacgcc tggcgatccc tgaacatgtc catcaggttc ttgcgaacct catcactcgt
     3721 tgcatcgacc ggtaatgcag gcaaattttg gtgtacggtc agtaaattgg acaccttcct
     3781 cttcttcttg ggcatggtcg aattcatcga tgatatcaga tctaattccg gcgcctagag
     3841 aaggagtgag ggctggataa agggaggatc gaggcggggt cgaacgagga ggttcaaggg
     3901 ggagagacgg ggcggatgga ggaagaggag gcggaggctt agggtgtaca aagggcttga
     3961 cccagggagg ggggtcaaaa gccaaggctt cccaggtcac gatgtagggg acctggtctg
     4021 ggtgtccatg cgggccaggt gaaaagacct tgatcttaac ctgggtgatg aggtctcggt
     4081 taaaggtgcc gtctcgcggc catccgacgt taaaggttgg ccattctgca gagcagaagg
     4141 taacccaacg tctcttcttg acatctaccg actggttgtg agcgatccgc tcgacatctt
     4201 tccagtgacc taaggtcaaa cttaagggag tggtaacagt ctggccctaa ttttcagaca
     4261 aatacagaaa cacagtcaga cagagacaac acagaacgat gctgcagcgc tgcagcagac
     4321 aagacgcgcg gcttcggttc caaaccgaaa gcaaaaattc agacggaggc gggaactgtt
     4381 ttaggttctc gtctcctacc agaaccacat atcctgacgg ggtcggattc cacatcgact
     4441 cccttcctca ggtcgggcca caaaaacggc ccccaaagtc cctgggacgt ctcccagggt
     4501 tgcggccggg tgttcagaac tcgtcagttc caccacgggt ccgccagata cagagctagt
     4561 tagctaacta gtacagacgc aggcgcaaac attagatgcc ggcacaaaca cgcacaaaga
     4621 cagagacaga cacgaaacga ccgctggcca gcttacctcc cggcgggggg gtcggtggtc
     4681 cctgggcagg ggtctccaaa tctcggtgga acctccaaat gaaagacccc cgaggtgggc
     4741 agtcaatcaa tctgaggaga ccctcccaag gatcagcgag tccacgattc ggatgcaaac
     4801 agcaagaggc tttattggga atacgggtac ccgggcgacg cagtctatcg gaggactggc
     4861 gcgccgagtg aggggttgtg ggctctttta ttgagctcgg ggagcagaag cgcgcgaaca
     4921 gaagcgagaa gcgaactgat tggttagttc aaataaggca cagggtcatt tcaggtcctt
     4981 ggggcaccct ggaaacatct gatggttctc tagaaactgc tgagggcggg accgcatctg
     5041 gggaccatct gttcttggcc ctgagccggg gcaggaactg cttaccacag atatcctgtt
     5101 tggcccatat tctgctgtct ctctgttcct aaccttgatc tgaacttctc tattctcagt
     5161 tatgtatttt ccatgccttg caaaatggcg ttacttaagc tagcttgcca aacctacagg
     5221 tggggtcttt cattcccccc tttttctgga gactaaataa aatcttttat tttatctatg
     5281 gctcgtactc tataggcttc agctggtgat attgttgagt caaaactaga gcctggacca
     5341 ctgatatcct gtctttaaca aattggacta atcgcctcta cgccggacgc atcgtggccg
     5401 gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc accgatgggg
     5461 aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt atggtggcag
     5521 gccccgtggc cgggggactg ttgggcgcca tctccttgca tgcaccattc cttgcgccgt
     5581 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     5641 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     5701 acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct
     5761 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
     5821 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     5881 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     5941 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     6001 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     6061 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     6121 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     6181 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     6241 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     6301 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     6361 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     6421 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     6481 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     6541 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     6601 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     6661 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     6721 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     6781 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     6841 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     6901 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     6961 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     7021 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     7081 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     7141 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     7201 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     7261 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     7321 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     7381 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     7441 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     7501 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     7561 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     7621 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     7681 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     7741 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     7801 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     7861 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     7921 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     7981 caaaccgcct ctccccgcgc gttggccgat tcattaatg
//
