LOCUS       sequence-245325-        6740 bp    DNA     circular SYN 14-APR-2025
DEFINITION  This plasmid expresses Bcl2 under the activation of tTA 
            trans-activator along with fluorescent marker tdTomato..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6740)
  AUTHORS   Wu Y, Ma L, Duyck K, Long CC, Moran A, Scheerer H, Blanck J, Peak A,
            Box A, Perera A, Yu CR
  TITLE     A Population of Navigator Neurons Is Essential for Olfactory Map 
            Formation during the Critical Period.
  JOURNAL   Neuron. 2018 Dec 5;100(5):1066-1082.e6. doi: 
            10.1016/j.neuron.2018.09.051. Epub 2018 Oct 25.
  PUBMED    30482691
REFERENCE   2  (bases 1 to 6740)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.neuron.2018.09.051"; journalName: "Neuron"; date: 
            "2018-12-5- 5"; volume: "100"; issue: "5"; pages: "1066-1082.e6"
COMMENT     Sequence Label: tetO-Bcl2-IRES-tdtomato(TET005)
FEATURES             Location/Qualifiers
     source          1..6740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..43
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     40..64
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          198..427
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     410..429
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      473..482
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1233..1784
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1377..1394)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             1808..3238
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGRYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     primer_bind     complement(3309..3326)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3315..3539
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3568..3588)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3568..3586)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3607..3623)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3607..3623)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3620..3642)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3631..3647
                     /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(3655..3661,3662..3679,3680..3685))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3700..3721
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3838..3855)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4009..4597)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4089..4108)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4768..5559,5560..5628))
                     /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     5391..5410
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5629..5733)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5759..6214)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5896..5917)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6108..6127
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6341..6363
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6355..6372
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6356..6372
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6382..6401
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6382..6400
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    join(6442..6460,6461..6483,6484..6502,6503..6525,6526..6544,
                     6545..6567,6568..6586,6587..6609,6610..6628,6629..6651,
                     6652..6670,6671..6693,6694..6712)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
ORIGIN
        1 tcgaggtagg cgtgtacggt gggaggccta tataagcaga gctcgtttag tgaaccgtca
       61 gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc
      121 cagcctccgc ggccccgaat tcaccacgtg gccgcatcga ttctagaatt cgctgtctgc
      181 gagggccagc tgttggggtg agtactccct ctcaaaagcg ggcatgactt ctgcgctaag
      241 attgtcagtt tccaaaaacg aggaggattt gatattcacc tggcccgcgg tgatgccttt
      301 gagggtggcc gcgtccatct ggtcagaaaa gacaatcttt ttgttgtcaa gcttgaggtg
      361 tggcaggctt gagatctggc catacacttg agtgacaatg acatccactt tgcctttctc
      421 tccacaggtg tccactccca ggtccaactg cagcccaagc ggatctacgc gtgccaccat
      481 ggcgcaagcc gggagaacag ggtatgataa ccgggagatc gtgatgaagt acatacatta
      541 taagctgtca cagaggggct acgagtggga tgctggagat gcggacgcgg cgcccctggg
      601 ggctgccccc acccctggca tcttctcctt ccagcctgag agcaacccaa tgcccgctgt
      661 gcaccgggac atggctgcca ggacgtctcc tctcaggccc ctcgttgcca ccgctgggcc
      721 tgcgctcagc cctgtgccac ctgtggtcca tctgaccctc cgccgggctg gggatgactt
      781 ctctcgtcgc taccgtcgtg acttcgcaga gatgtccagt cagctgcacc tgacgccctt
      841 caccgcgagg ggacgctttg ccacggtggt ggaggaactc ttcagggatg gggtgaactg
      901 ggggaggatt gtggccttct ttgagttcgg tggggtcatg tgtgtggaga gcgtcaacag
      961 ggagatgtca cccctggtgg acaacatcgc cctgtggatg actgagtacc tgaaccggca
     1021 tctgcacacc tggatccagg ataacggagg ctgggatgcc tttgtggaac tatatggccc
     1081 cagcatgcga cctctgtttg atttctcctg gctgtctctg aagaccctgc tcagcctggc
     1141 cctggtcggg gcctgcatca ctctgggtgc atacctgggc cacaagtgat taattaagtt
     1201 caagcttcga attccgccct cccccccccc taacgttact ggccgaagcc gcttggaata
     1261 aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt
     1321 gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct
     1381 cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc
     1441 ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga
     1501 caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc
     1561 ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctaagcgta
     1621 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg
     1681 cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg
     1741 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca caagctagca
     1801 tccgatgatg gtgagcaagg gcgaggaggt catcaaagag ttcatgcgct tcaaggtgcg
     1861 catggagggc tccatgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     1921 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc
     1981 ctgggacatc ctgtcccccc agttcatgta cggctccaag gcgtacgtga agcaccccgc
     2041 cgacatcccc gattacaaga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     2101 gaacttcgag gacggcggtc tggtgaccgt gacccaggac tcctccctgc aggacggcac
     2161 gctgatctac aaggtgaaga tgcgcggcac caacttcccc cccgacggcc ccgtaatgca
     2221 gaagaagacc atgggctggg aggcctccac cgagcgcctg tacccccgcg acggcgtgct
     2281 gaagggcgag atccaccagg ccctgaagct gaaggacggc ggccactacc tggtggagtt
     2341 caagaccatc tacatggcca agaagcccgt gcaactgccc ggctactact acgtggacac
     2401 caagctggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgagcgctc
     2461 cgagggccgc caccacctgt tcctggggca tggcaccggc agcaccggca gcggcagctc
     2521 cggcaccgcc tcctccgagg acaacaacat ggccgtcatc aaagagttca tgcgcttcaa
     2581 ggtgcgcatg gagggctcca tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     2641 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc
     2701 cttcgcctgg gacatcctgt ccccccagtt catgtacggc tccaaggcgt acgtgaagca
     2761 ccccgccgac atccccgatt acaagaagct gtccttcccc gagggcttca agtgggagcg
     2821 cgtgatgaac ttcgaggacg gcggtctggt gaccgtgacc caggactcct ccctgcagga
     2881 cggcacgctg atctacaagg tgaagatgcg cggcaccaac ttcccccccg acggccccgt
     2941 aatgcagaag aagaccatgg gctgggaggc ctccaccgag cgcctgtacc cccgcgacgg
     3001 cgtgctgaag ggcgagatcc accaggccct gaagctgaag gacggcggcc gctacctggt
     3061 ggagttcaag accatctaca tggccaagaa gcccgtgcaa ctgcccggct actactacgt
     3121 ggacaccaag ctggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     3181 gcgctccgag ggccgccacc acctgttcct gtacggcatg gacgagctgt acaagtaaga
     3241 attctgcaga tatccagcac agtggcggcc gctcgagtct agagggcccg tttaaacccg
     3301 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3361 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3421 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3481 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     3541 ttggccggcc agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
     3601 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     3661 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     3721 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     3781 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     3841 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3901 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3961 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4021 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4081 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4141 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4201 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4261 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4321 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4381 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4441 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4501 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4561 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4621 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4681 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     4741 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4801 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4861 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     4921 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4981 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5041 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5101 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5161 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5221 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5281 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5341 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5401 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     5461 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     5521 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     5581 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     5641 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     5701 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa
     5761 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     5821 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     5881 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     5941 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     6001 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     6061 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     6121 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     6181 cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac
     6241 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     6301 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     6361 acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattggg tacggcgcgc
     6421 cggatccctc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac
     6481 cactccctat cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata
     6541 gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga
     6601 gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca
     6661 gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa
     6721 agtcgagctc ggtacccggg
//
