LOCUS       sequence-430613-        6064 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construct 3: TetR - Lipid binding domain KR-phi - TetR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6064)
  AUTHORS   Park J, Berthoux C, Hoyos-Ramirez E, Shan L, Morimoto-Tomita M, Wang
            Y, Castillo PE, Tomita S
  TITLE     Chemogenetic regulation of the TARP-lipid interaction mimics LTP and
            reversibly modifies behavior.
  JOURNAL   Cell Rep. 2023 Aug 29;42(8):112826. doi: 
            10.1016/j.celrep.2023.112826. Epub 2023 Jul 19.
  PUBMED    37471228
REFERENCE   2  (bases 1 to 6064)
  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: "Cell Rep.";
            date: "2023-08-29"; volume: "42(8):112826. doi"; pages: " 
            10.1016/j.celrep.2023.112826. Epub 2023 Jul 19"
COMMENT     Sequence Label: pEGFP-C2-TetR-KRf-TetR
FEATURES             Location/Qualifiers
     source          1..6064
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        80..271
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     228..248
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(273..377)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      404..859
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(491..510)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     701..722
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    866..987
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(877..896)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     931..950
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1175..1795)
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     CDS             complement(1847..2467)
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     CDS             complement(join(2510..3220,3221..3223,3224..3226))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(2510..3220,3221..3223,3224..3226))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2552..2573)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2901..2920
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3160..3181
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3223..3232
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3271..3474)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3300..3320)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3475..3778
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3953..4541)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4033..4052)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4870..4917
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4942..4961
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5149..5943)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(5149..5943)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5260..5279)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5870..5889
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5978..6064,1..271))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5992..6064,1..63)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg
       61 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat
      121 acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg
      181 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca
      241 cacattccac agctggttct ttccgcctca ggactcttcc tttttcaata ttattgaagc
      301 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
      361 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc
      421 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
      481 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
      541 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
      601 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
      661 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
      721 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
      781 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
      841 atattaacgc ttacaattta cgcgttaaga tacattgatg agtttggaca aaccacaact
      901 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
      961 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     1021 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg
     1081 gctgattatg atcagttatc tagatccggt ggatcccggg cccgcggtac cgtcgactgc
     1141 agaattccta cacgggcgtg gttttcctgt tgagagaccc actttcacat ttaagttgtt
     1201 tttctaatcc gcatatgatc aattcaaggc cgaataagaa ggctggctct gcaccttggt
     1261 gatcaaataa ttcgatagct tgtcgtaata atggcggcat actatcagta gtaggtgttt
     1321 ccctttcttc tttagcgact tgatgctctt gatcttccaa tacgcaacct aaagtaaaat
     1381 gccccacagc gctgagtgca tataatgcat tctctagtga aaaaccttgt tggcataaaa
     1441 aggctaattg attttcgaga gtttcatact gtttttctgt aggccgtgta cctaaatgta
     1501 cttttgctcc atcgcgatga cttagtaaag cacatctaaa acttttagcg ttattacgta
     1561 aaaaatcttg ccagctttcc ccttctaaag ggcaaaagtg agtatggtgc ctatctaaca
     1621 tctcaatggc taaggcgtcg agcaaagccc gcttattttt tacatgccaa tacaatgtag
     1681 gctgctctac acctagcttc tgggcgagtt tacgggttgt taaaccttcg attccgacct
     1741 cattaagcag ctctaatgcg ctgttaatca ctttactttt atctaatcta gacatgcttt
     1801 tcagcttccg gaggaatttt ctcagcctct tccaaaattt cttgccagac ccactttcac
     1861 atttaagttg tttttctaat ccgcatatga tcaattcaag gccgaataag aaggctggct
     1921 ctgcaccttg gtgatcaaat aattcgatag cttgtcgtaa taatggcggc atactatcag
     1981 tagtaggtgt ttccctttct tctttagcga cttgatgctc ttgatcttcc aatacgcaac
     2041 ctaaagtaaa atgccccaca gcgctgagtg catataatgc attctctagt gaaaaacctt
     2101 gttggcataa aaaggctaat tgattttcga gagtttcata ctgtttttct gtaggccgtg
     2161 tacctaaatg tacttttgct ccatcgcgat gacttagtaa agcacatcta aaacttttag
     2221 cgttattacg taaaaaatct tgccagcttt ccccttctaa agggcaaaag tgagtatggt
     2281 gcctatctaa catctcaatg gctaaggcgt cgagcaaagc ccgcttattt tttacatgcc
     2341 aatacaatgt aggctgctct acacctagct tctgggcgag tttacgggtt gttaaacctt
     2401 cgattccgac ctcattaagc agctctaatg cgctgttaat cactttactt ttatctaatc
     2461 tagacattgc ctcgcggctg cgctcgctcg agatctgagt ccggccggac ttgtacagct
     2521 cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc
     2581 gcttctcgtt ggggtctttg ctcagggcgg actgggtgct caggtagtgg ttgtcgggca
     2641 gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg agctgcacgc
     2701 tgccgtcctc gatgttgtgg cggatcttga agttcacctt gatgccgttc ttctgcttgt
     2761 cggccatgat atagacgttg tggctgttgt agttgtactc cagcttgtgc cccaggatgt
     2821 tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg gtgtcgccct
     2881 cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct
     2941 cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc atgtggtcgg
     3001 ggtagcggct gaagcactgc acgccgtagg tcagggtggt cacgagggtg ggccagggca
     3061 cgggcagctt gccggtggtg cagatgaact tcagggtcag cttgccgtag gtggcatcgc
     3121 cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca
     3181 ggatgggcac caccccggtg aacagctcct cgcccttgct caccatggtg gcgaccggta
     3241 gcgctagcgg atctgacggt tcactaaacc agctctgctt atatagacct cccaccgtac
     3301 acgcctaccg cccatttgcg tcaatggggc ggagttgtta cgacattttg gaaagtcccg
     3361 ttgattttgg tgccaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc
     3421 ccgtgagtca aaccgctatc cacgcccatt gatgtactgc caaaaccgca tcaccatggt
     3481 aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta
     3541 ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg
     3601 catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacgcattg
     3661 acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa
     3721 tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcg
     3781 gaactccata tatgggctat gaactaatga ccccgtaatt gattactatt aataactaat
     3841 gcatggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3901 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3961 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4021 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4081 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4141 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4201 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4261 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4321 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4381 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4441 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4501 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4561 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4621 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4681 aaagtatata tgagtaacct gaggctatgg cagggcctgc cgccccgacg ttggctgcga
     4741 gccctgggcc ttcacccgaa cttggggggt ggggtgggga aaaggaagaa acgcgggcgt
     4801 attggcccca atggggtctc ggtggggtat cgacagagtg ccagccctgg gaccgaaccc
     4861 cgcgtttatg aacaaacgac ccaacaccgt gcgttttatt ctgtcttttt attgccgtca
     4921 tagcgcgggt tccttccggt attgtctcct tccgtgtttc agttagcctc cccctagggt
     4981 gggcgaagaa ctccagcatg agatccccgc gctggaggat catccagccg gcgtcccgga
     5041 aaacgattcc gaagcccaac ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg
     5101 gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc agaagaactc
     5161 gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac
     5221 gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc
     5281 tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg
     5341 gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc
     5401 gccgtcgggc atgctcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg
     5461 ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc
     5521 gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg
     5581 ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag
     5641 atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc
     5701 gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc
     5761 ttgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg
     5821 cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata
     5881 gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat
     5941 catgcgaaac gatcctcatc ctgtctcttg atcgatcttt gcaaaagcct aggcctccaa
     6001 aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat
     6061 aaat
//
