LOCUS       sequence-316176-        7952 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid expresses Fzd1 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 7952)
  AUTHORS   Wu Y, Ma L, Qiu Q, Xu W, Misra A, Duyck K, Blanck J, Scott AR, Chen 
            S, Hassan H, Corbin TJ, Moran A, Hall K, Li H, Perera A, Yu CR
  TITLE     Molecular Control of Circuit Plasticity and the Permanence of 
            Imprinted Odor Memory
  JOURNAL   bioRxiv 2022
REFERENCE   2  (bases 1 to 7952)
  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: "bioRxiv 
            2022"
COMMENT     Sequence Label: tetO-Fzd1-IRES-tdTomato (TET006)
FEATURES             Location/Qualifiers
     source          1..7952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     12..34
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     26..43
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     27..43
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     53..72
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        53..71
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    join(113..131,132..154,155..173,174..196,197..215,216..238,
                     239..257,258..280,281..299,300..322,323..341,342..364,
                     365..383)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        416..454
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     451..475
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          609..838
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     821..840
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    2856..3407
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3000..3017)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             3431..4861
                     /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(4932..4949)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4938..5162
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5191..5211)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5191..5209)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5230..5246)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5230..5246)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5243..5265)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5254..5270
                     /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(5278..5284,5285..5302,5303..5308))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5323..5344
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5461..5478)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5632..6220)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5712..5731)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6391..7182,7183..7251))
                     /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     7014..7033
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7252..7356)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7382..7837)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7519..7540)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7731..7750
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga
       61 ctcactatag ggcgaattgg gtacggcgcg ccggatccct cgagtttacc actccctatc
      121 agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag agaaaagtga
      181 aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc
      241 cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa
      301 aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta
      361 ccactcccta tcagtgatag agaaaagtga aagtcgagct cggtacccgg gtcgaggtag
      421 gcgtgtacgg tgggaggcct atataagcag agctcgttta gtgaaccgtc agatcgcctg
      481 gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg
      541 cggccccgaa ttcaccacgt ggccgcatcg attctagaat tcgctgtctg cgagggccag
      601 ctgttggggt gagtactccc tctcaaaagc gggcatgact tctgcgctaa gattgtcagt
      661 ttccaaaaac gaggaggatt tgatattcac ctggcccgcg gtgatgcctt tgagggtggc
      721 cgcgtccatc tggtcagaaa agacaatctt tttgttgtca agcttgaggt gtggcaggct
      781 tgagatctgg ccatacactt gagtgacaat gacatccact ttgcctttct ctccacaggt
      841 gtccactccc aggtccaact gcagcccaag cggatctacg cgtatggctg aggaggcggc
      901 gcctagcgag tcccgggccg ccggccggct gagcttggaa ctttgtgccg aagcactccc
      961 gggccggcgg gaggaggtgg ggcacgagga cacggccagc caccgccgcc cccgggctga
     1021 tccccggcgt tgggctagcg ggctgctgct gctgctttgg ttgctggagg ctcctctgct
     1081 tttgggggtc cgagcgcagg cggcgggcca ggtatccggg ccgggccagc aagccccgcc
     1141 gccgccccag ccccagcaga gcgggcagca gtacaacggc gaacggggca tctccatccc
     1201 ggaccacggc tactgccagc ccatctccat cccgctgtgc acggacatcg cgtacaacca
     1261 gaccatcatg cccaacctgc tgggccacac gaatcaggag gacgccggtc tggaggtgca
     1321 ccagttctac cctctggtga aggtgcagtg ctccgccgag ctcaagttct tcctgtgctc
     1381 catgtacgcg cctgtgtgca ccgtactgga gcaggcgcta ccgccctgcc gctccctgtg
     1441 cgagcgcgca cgccagggct gcgaggcgct catgaacaag ttcggcttcc agtggccaga
     1501 cacactcaag tgcgagaagt tcccggtgca cggcgcagga gagctgtgcg tgggccagaa
     1561 cacgtccgac aaaggcaccc caactccctc cttgctacca gagttctgga ccagtaatcc
     1621 gcagcacggc ggcggtggtt accgcggcgg ctacccgggg ggtgccggga cggtggagcg
     1681 gggaaagttc tcctgcccgc gcgccctcag ggtgccctcc tacctcaact accactttct
     1741 gggggagaag gactgcggcg caccctgcga acccaccaag gtttacgggc tcatgtactt
     1801 cgggccagag gagctgcgct tctcgcgcac ctggataggc atctggtccg tgctgtgctg
     1861 cgcctccacg ctcttcacgg tgctcacgta cctagtggac atgcggcgct tcagctaccc
     1921 ggaacggccc atcattttcc tgtccggctg ttacacagcg gtggcggtgg cctacatcgc
     1981 tggctttctg ttggaggacc gggtggtgtg caacgacaag tttgcagagg acggggcgcg
     2041 cacggtggcg cagggcacta agaaagaagg ctgcactata ctctttatga tgctctactt
     2101 cttcagcatg gccagctcca tctggtgggt gatcctgtcc ctcacctggt tcctggcagc
     2161 cggcatgaag tggggccacg aagccatcga ggccaactca cagtatttcc atttagccgc
     2221 ctgggctgtg ccagccatca aaactataac catcttggcg ttgggccagg tggatggcga
     2281 cgtactgagc ggagtgtgtt ttgtggggct caacaacgtg gacgcactgc gtggctttgt
     2341 gctggcgcct ctcttcgttt atctgttcat tggcacttct ttcctgctgg ccggtttcgt
     2401 gtcactcttc cgcatccgca ccatcatgaa gcatgacggc accaagacag agaagctgga
     2461 gaagctcatg gtgcgcatcg gagtcttcag tgtcctctac actgtgccgg ccaccatcgt
     2521 catcgcctgc tacttctatg aacaggcctt tcgggaccag tgggagcgca gctgggtggc
     2581 ccagagctgc aagagttatg ccatcccttg ccctcacctc cagggaggtg gaggagtccc
     2641 accacacccg cccatgagcc cagactttac agtcttcatg atcaagtatc tcatgacgct
     2701 gattgtgggc atcacatcgg gcttctggat ctggtccggc aagacactga attcctggag
     2761 gaagttctac acgaggctta ccaacagcaa acagggggag actaccgtct gattaattaa
     2821 gttcaagctt cgaattccgc cctccccccc ccctaacgtt actggccgaa gccgcttgga
     2881 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa
     2941 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
     3001 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc
     3061 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
     3121 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca
     3181 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctaagc
     3241 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg
     3301 gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc
     3361 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaagcta
     3421 gcatccgatg atggtgagca agggcgagga ggtcatcaaa gagttcatgc gcttcaaggt
     3481 gcgcatggag ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg
     3541 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggcggcc ccctgccctt
     3601 cgcctgggac atcctgtccc cccagttcat gtacggctcc aaggcgtacg tgaagcaccc
     3661 cgccgacatc cccgattaca agaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     3721 gatgaacttc gaggacggcg gtctggtgac cgtgacccag gactcctccc tgcaggacgg
     3781 cacgctgatc tacaaggtga agatgcgcgg caccaacttc ccccccgacg gccccgtaat
     3841 gcagaagaag accatgggct gggaggcctc caccgagcgc ctgtaccccc gcgacggcgt
     3901 gctgaagggc gagatccacc aggccctgaa gctgaaggac ggcggccact acctggtgga
     3961 gttcaagacc atctacatgg ccaagaagcc cgtgcaactg cccggctact actacgtgga
     4021 caccaagctg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgagcg
     4081 ctccgagggc cgccaccacc tgttcctggg gcatggcacc ggcagcaccg gcagcggcag
     4141 ctccggcacc gcctcctccg aggacaacaa catggccgtc atcaaagagt tcatgcgctt
     4201 caaggtgcgc atggagggct ccatgaacgg ccacgagttc gagatcgagg gcgagggcga
     4261 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gcggccccct
     4321 gcccttcgcc tgggacatcc tgtcccccca gttcatgtac ggctccaagg cgtacgtgaa
     4381 gcaccccgcc gacatccccg attacaagaa gctgtccttc cccgagggct tcaagtggga
     4441 gcgcgtgatg aacttcgagg acggcggtct ggtgaccgtg acccaggact cctccctgca
     4501 ggacggcacg ctgatctaca aggtgaagat gcgcggcacc aacttccccc ccgacggccc
     4561 cgtaatgcag aagaagacca tgggctggga ggcctccacc gagcgcctgt acccccgcga
     4621 cggcgtgctg aagggcgaga tccaccaggc cctgaagctg aaggacggcg gccgctacct
     4681 ggtggagttc aagaccatct acatggccaa gaagcccgtg caactgcccg gctactacta
     4741 cgtggacacc aagctggaca tcacctccca caacgaggac tacaccatcg tggaacagta
     4801 cgagcgctcc gagggccgcc accacctgtt cctgtacggc atggacgagc tgtacaagta
     4861 agaattctgc agatatccag cacagtggcg gccgctcgag tctagagggc ccgtttaaac
     4921 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     4981 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     5041 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     5101 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     5161 ggcttggccg gccagctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc
     5221 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5281 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5341 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5401 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5461 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5521 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5581 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5641 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5701 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5761 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5821 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5881 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5941 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6001 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6061 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6121 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6181 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6241 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6301 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6361 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6421 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6481 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6541 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6601 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6661 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6721 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6781 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6841 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6901 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6961 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7021 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7081 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7141 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7201 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7261 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7321 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7381 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     7441 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     7501 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     7561 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     7621 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     7681 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     7741 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     7801 acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc
     7861 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     7921 ggatgtgctg caaggcgatt aagttgggta ac
//
