LOCUS       sequence-138554-        7338 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of Cre.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7338)
  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 7338)
  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: pMSCVpuro-Cre
FEATURES             Location/Qualifiers
     source          1..7338
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..517
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    581..922
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             989..1405
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1335..1357
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1373..1389
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1443..1463
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1464..2492
                     /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"
     promoter        2514..3013
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2549..2572)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2946..2965
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3034..3633
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3034..3053)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3530..3550
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3733..4247
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4409..4425
                     /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)."
     promoter        complement(join(4433..4439,4440..4457,4458..4463))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4478..4499
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4616..4633)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4787..5375)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4867..4886)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5546..6337,6338..6406))
                     /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     6169..6188
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6407..6511)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6579..6597
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6635..6657)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6757..6776
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6970..6992
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7120..7139
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
      121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
      241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
      301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
      421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
      481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
      541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
      601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
      661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
      721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
      781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
      841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
      901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
      961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
     1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
     1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
     1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
     1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
     1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
     1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
     1381 agccctcact ccttctctag gcgccggaat tagatctgat atcatcgatg aattcgacca
     1441 tgcccaagaa gaagaggaag gtgtccaatt tactgaccgt acaccaaaat ttgcctgcat
     1501 taccggtcga tgcaacgagt gatgaggttc gcaagaacct gatggacatg ttcagggatc
     1561 gccaggcgtt ttctgagcat acctggaaaa tgcttctgtc cgtttgccgg tcgtgggcgg
     1621 catggtgcaa gttgaataac cggaaatggt ttcccgcaga acctgaagat gttcgcgatt
     1681 atcttctata tcttcaggcg cgcggtctgg cagtaaaaac tatccagcaa catttgggcc
     1741 agctaaacat gcttcatcgt cggtccgggc tgccacgacc aagtgacagc aatgctgttt
     1801 cactggttat gcggcggatc cgaaaagaaa acgttgatgc cggtgaacgt gcaaaacagg
     1861 ctctagcgtt cgaacgcact gatttcgacc aggttcgttc actcatggaa aatagcgatc
     1921 gctgccagga tatacgtaat ctggcatttc tggggattgc ttataacacc ctgttacgta
     1981 tagccgaaat tgccaggatc agggttaaag atatctcacg tactgacggt gggagaatgt
     2041 taatccatat tggcagaacg aaaacgctgg ttagcaccgc aggtgtagag aaggcactta
     2101 gcctgggggt aactaaactg gtcgagcgat ggatttccgt ctctggtgta gctgatgatc
     2161 cgaataacta cctgttttgc cgggtcagaa aaaatggtgt tgccgcgcca tctgccacca
     2221 gccagctatc aactcgcgcc ctggaaggga tttttgaagc aactcatcga ttgatttacg
     2281 gcgctaagga tgactctggt cagagatacc tggcctggtc tggacacagt gcccgtgtcg
     2341 gagccgcgcg agatatggcc cgcgctggag tttcaatacc ggagatcatg caagctggtg
     2401 gctggaccaa tgtaaatatt gtcatgaact atatccgtaa cctggatagt gaaacagggg
     2461 caatggtgcg cctgctggaa gatggcgatt agtcgaggtt aacgaattct accgggtagg
     2521 ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg
     2581 gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc
     2641 ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt
     2701 cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac
     2761 tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag
     2821 cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc
     2881 cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt cctccggagg
     2941 cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc
     3001 tccgggcctt tcgacctgca gcccaagctt accatgaccg agtacaagcc cacggtgcgc
     3061 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     3121 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     3181 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     3241 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     3301 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     3361 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     3421 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     3481 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     3541 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     3601 acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg
     3661 accgaaagga gcgcacgacc ccatgcatcg ataaaataaa agattttatt tagtctccag
     3721 aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca
     3781 ttttgcaagg catggaaaat acataactga gaatagagaa gttcagatca aggttaggaa
     3841 cagagagaca gcagaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
     3901 ctcagggcca agaacagatg gtccccagat gcggtcccgc cctcagcagt ttctagagaa
     3961 ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact
     4021 aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa
     4081 gagcccacaa cccctcactc ggcgcgccag tcctccgata gactgcgtcg cccgggtacc
     4141 cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg
     4201 agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcatggg taacagtttc
     4261 ttgaagttgg agaacaacat tctgagggta ggagtcgaat attaagtaat cctgactcaa
     4321 ttagccactg ttttgaatcc acatactcca atactcctga aatagttcat tatggacagc
     4381 gcagaaagag ctggggagaa ttgtgaaatt gttatccgct cacaattcca cacaacatac
     4441 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     4501 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     4561 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     4621 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4681 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4741 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4801 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4861 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4921 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4981 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5041 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5101 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5161 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5221 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5281 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5341 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5401 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5461 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5521 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5581 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5641 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5701 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5761 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5821 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5881 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5941 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6001 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6061 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6121 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6181 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6241 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6301 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6361 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6421 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6481 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     6541 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     6601 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     6661 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     6721 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     6781 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     6841 ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     6901 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     6961 cagggttttc ccagtcacga cgttgtaaaa cgacggcgca aggaatggtg catgcaagga
     7021 gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc
     7081 gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc
     7141 gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggcg
     7201 attagtccaa tttgttaaag acaggatatc agtggtccag gctctagttt tgactcaaca
     7261 atatcaccag ctgaagccta tagagtacga gccatagata aaataaaaga ttttatttag
     7321 tctccagaaa aagggggg
//
