LOCUS       40924_7981        6569 bp DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression of Cre recombinase fused to GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6569)
  AUTHORS   Matsuda T, Cepko CL
  TITLE     Controlled expression of transgenes introduced by in vivo 
            electroporation.
  JOURNAL   Proc Natl Acad Sci U S A. 2007 Jan 5. ():.
  PUBMED    17209010
REFERENCE   2  (bases 1 to 6569)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6569)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A. 2007 Jan 5. ():."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6569
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1674
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and
                     rabbit beta-globin"
     primer_bind     1682..1701
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids,
                     forward primer"
     CDS             1741..2769
                     /codon_start=1
                     /label=Cre
                     /note="site-specific recombinase"
                     /translation="MANLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     CDS             2785..3501
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3591..3610)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3656..3711
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3710..3729)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also
                     called rb-glob-pA-term-R"
     primer_bind     complement(4072..4088)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4096..4112)
                     /label=lac operator
                     /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(4120..4150)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4165..4186)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4244..4440
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     polyA_signal    4446..4580
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4648..4665)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4819..5407)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5581..6438)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6439..6543)
                     /label=AmpR promoter
     promoter        6569
                     /label=CAG
                     /note="CMV early enhancer fused to modified chicken 
                     beta-actin promoter"
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
     1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1621 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1681 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattctg agccgccacc
     1741 atggccaatt tactgaccgt acaccaaaat ttgcctgcat taccggtcga tgcaacgagt
     1801 gatgaggttc gcaagaacct gatggacatg ttcagggatc gccaggcgtt ttctgagcat
     1861 acctggaaaa tgcttctgtc cgtttgccgg tcgtgggcgg catggtgcaa gttgaataac
     1921 cggaaatggt ttcccgcaga acctgaagat gttcgcgatt atcttctata tcttcaggcg
     1981 cgcggtctgg cagtaaaaac tatccagcaa catttgggcc agctaaacat gcttcatcgt
     2041 cggtccgggc tgccacgacc aagtgacagc aatgctgttt cactggttat gcggcggatc
     2101 cgaaaagaaa acgttgatgc cggtgaacgt gcaaaacagg ctctagcgtt cgaacgcact
     2161 gatttcgacc aggttcgttc actcatggaa aatagcgatc gctgccagga tatacgtaat
     2221 ctggcatttc tggggattgc ttataacacc ctgttacgta tagccgaaat tgccaggatc
     2281 agggttaaag atatctcacg tactgacggt gggagaatgt taatccatat tggcagaacg
     2341 aaaacgctgg ttagcaccgc aggtgtagag aaggcactta gcctgggggt aactaaactg
     2401 gtcgagcgat ggatttccgt ctctggtgta gctgatgatc cgaataacta cctgttttgc
     2461 cgggtcagaa aaaatggtgt tgccgcgcca tctgccacca gccagctatc aactcgcgcc
     2521 ctggaaggga tttttgaagc aactcatcga ttgatttacg gcgctaagga tgactctggt
     2581 cagagatacc tggcctggtc tggacacagt gcccgtgtcg gagccgcgcg agatatggcc
     2641 cgcgctggag tttcaatacc ggagatcatg caagctggtg gctggaccaa tgtaaatatt
     2701 gtcatgaact atatccgtaa cctggatagt gaaacagggg caatggtgcg cctgctggaa
     2761 gatggcgatg gaccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
     2821 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     2881 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     2941 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     3001 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     3061 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     3121 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     3181 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     3241 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     3301 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     3361 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     3421 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     3481 ggcatggacg agctgtacaa gtaaagcggc cgcactcctc aggtgcaggc tgcctatcag
     3541 aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc
     3601 tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa
     3661 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
     3721 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca
     3781 acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat
     3841 atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag
     3901 atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta
     3961 catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct
     4021 cttctcttat ggagatccct cgacctgcag cccaagcttg gcgtaatcat ggtcatagct
     4081 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4141 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4201 actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct caattagtca
     4261 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4321 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     4381 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4441 aagctaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4501 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4561 tatcttatca tgtctggatc cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4621 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4681 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4741 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4801 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4861 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4921 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4981 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5041 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5101 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5161 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5221 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5281 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5341 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5401 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5461 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5521 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5581 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5641 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5701 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5761 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5821 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5881 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5941 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6001 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6061 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6121 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6181 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6241 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6301 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6361 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6421 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6481 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6541 gcgcacattt ccccgaaaag tgccacctg
//
