LOCUS       Exported                5893 bp ds-DNA     circular SYN 26-JUL-2023
DEFINITION  Mammalian expression of conditionally active form of Cre recombinase
            (not as tightly regulated as pCAG-ERT2CreERT2).
ACCESSION   .
VERSION     .
KEYWORDS    pCAG-CreERT2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5893)
  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 5893)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 26, 2023 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5893
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     309..331
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     393..412
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      446..455
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             452..1480
                     /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="MANLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     CDS             1493..2428
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     primer_bind     complement(2522..2541)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2587..2642
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2641..2660)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3003..3019)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3003..3019)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3016..3038)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3027..3043
                     /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(3051..3081)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3096..3117
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3176..3371
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3222..3357
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3284..3303
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3377..3511
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3414..3433)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3468..3487
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3579..3596)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3750..4338)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3830..3849)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4509..5369)
                     /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     5132..5151
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5370..5474)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5505..5884
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg
       61 ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctgtgcgga
      121 gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga agcggtgcgg
      181 cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct
      241 tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg
      301 cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt
      361 tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg tgctgtctca
      421 tcattttggc aaagaattct gagccgccac catggccaat ttactgaccg tacaccaaaa
      481 tttgcctgca ttaccggtcg atgcaacgag tgatgaggtt cgcaagaacc tgatggacat
      541 gttcagggat cgccaggcgt tttctgagca tacctggaaa atgcttctgt ccgtttgccg
      601 gtcgtgggcg gcatggtgca agttgaataa ccggaaatgg tttcccgcag aacctgaaga
      661 tgttcgcgat tatcttctat atcttcaggc gcgcggtctg gcagtaaaaa ctatccagca
      721 acatttgggc cagctaaaca tgcttcatcg tcggtccggg ctgccacgac caagtgacag
      781 caatgctgtt tcactggtta tgcggcggat ccgaaaagaa aacgttgatg ccggtgaacg
      841 tgcaaaacag gctctagcgt tcgaacgcac tgatttcgac caggttcgtt cactcatgga
      901 aaatagcgat cgctgccagg atatacgtaa tctggcattt ctggggattg cttataacac
      961 cctgttacgt atagccgaaa ttgccaggat cagggttaaa gatatctcac gtactgacgg
     1021 tgggagaatg ttaatccata ttggcagaac gaaaacgctg gttagcaccg caggtgtaga
     1081 gaaggcactt agcctggggg taactaaact ggtcgagcga tggatttccg tctctggtgt
     1141 agctgatgat ccgaataact acctgttttg ccgggtcaga aaaaatggtg ttgccgcgcc
     1201 atctgccacc agccagctat caactcgcgc cctggaaggg atttttgaag caactcatcg
     1261 attgatttac ggcgctaagg atgactctgg tcagagatac ctggcctggt ctggacacag
     1321 tgcccgtgtc ggagccgcgc gagatatggc ccgcgctgga gtttcaatac cggagatcat
     1381 gcaagctggt ggctggacca atgtaaatat tgtcatgaac tatatccgta acctggatag
     1441 tgaaacaggg gcaatggtgc gcctgctgga agatggcgat ctcgagccat ctgctggaga
     1501 catgagagct gccaaccttt ggccaagccc gctcatgatc aaacgctcta agaagaacag
     1561 cctggccttg tccctgacgg ccgaccagat ggtcagtgcc ttgttggatg ctgagccccc
     1621 catactctat tccgagtatg atcctaccag acccttcagt gaagcttcga tgatgggctt
     1681 actgaccaac ctggcagaca gggagctggt tcacatgatc aactgggcga agagggtgcc
     1741 aggctttgtg gatttgaccc tccatgatca ggtccacctt ctagaatgtg cctggctaga
     1801 gatcctgatg attggtctcg tctggcgctc catggagcac ccagtgaagc tactgtttgc
     1861 tcctaacttg ctcttggaca ggaaccaggg aaaatgtgta gagggcatgg tggagatctt
     1921 cgacatgctg ctggctacat catctcggtt ccgcatgatg aatctgcagg gagaggagtt
     1981 tgtgtgcctc aaatctatta ttttgcttaa ttctggagtg tacacatttc tgtccagcac
     2041 cctgaagtct ctggaagaga aggaccatat ccaccgagtc ctggacaaga tcacagacac
     2101 tttgatccac ctgatggcca aggcaggcct gaccctgcag cagcagcacc agcggctggc
     2161 ccagctcctc ctcatcctct cccacatcag gcacatgagt aacaaaggca tggagcatct
     2221 gtacagcatg aagtgcaaga acgtggtgcc cctctatgac ctgctgctgg aggcggcgga
     2281 cgcccaccgc ctacatgcgc ccactagccg tggaggggca tccgtggagg agacggacca
     2341 aagccacttg gccactgcgg gctctacttc atcgcattcc ttgcaaaagt attacatcac
     2401 gggggaggca gagggtttcc ctgccacagc ttgatagcgg ccgcactcct caggtgcagg
     2461 ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag
     2521 atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt
     2581 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct
     2641 cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt
     2701 agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag
     2761 gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga
     2821 cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa
     2881 aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca
     2941 tagctgtccc tcttctctta tggagatccc tcgacctgca gcccaagctt ggcgtaatca
     3001 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     3061 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     3121 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc
     3181 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3241 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3301 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3361 gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     3421 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     3481 actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg
     3541 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3601 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3661 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3721 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3781 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3841 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3901 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3961 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4021 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4081 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4141 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4201 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4261 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4321 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4381 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4441 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4501 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4561 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4621 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4681 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4741 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4801 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4861 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4921 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4981 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5041 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5101 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5161 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5221 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5281 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5341 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5401 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5461 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg ggtcgacatt gattattgac
     5521 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     5581 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     5641 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     5701 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     5761 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     5821 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     5881 catggtcgag gtg
//
