LOCUS       sequence-7335-ex        7218 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7218)
  AUTHORS   Jiang W, Zhou L, Breyer B, Feng T, Cheng H, Haydon R, Ishikawa A, He
            TC
  TITLE     Tetracycline-regulated gene expression mediated by a novel chimeric 
            repressor that recruits histone deacetylases in mammalian cells.
  JOURNAL   J Biol Chem. 2001 Nov 30. 276(48):45168-74.
  PUBMED    11581265
REFERENCE   2  (bases 1 to 7218)
  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: "J Biol 
            Chem. 2001 Nov 30. 276(48):45168-74."
COMMENT     Sequence Label: pTHE
FEATURES             Location/Qualifiers
     source          1..7218
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        262..641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        642..845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        890..908
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          948..1520
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     1528..1547
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     promoter        1575..1593
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1604..2224
                     /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"
     primer_bind     complement(1677..1696)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     misc_feature    2378..2950
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2401..2952
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2545..2562)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2772..2791
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3019..3813
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVLRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3073..3092)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3683..3702
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3987..4121
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4024..4043)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4078..4097
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             4130..4171
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     enhancer        4176..4555
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4556..4759
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    4761..4779
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4782..4800
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4803..4821
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4824..4842
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(4977..4994)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4983..5207
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5295..5312)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5466..6051)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5546..5565)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6222..7013,7014..7082))
                     /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     6845..6864
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7083..7187)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattaatta aggcgcgccg
      181 tttaaactaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      241 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      661 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      781 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      841 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      901 actataggga gacccaagct ggctagatcg atcctgagaa cttcagggtg agtttgggga
      961 cccttgattg ttctttcttt ttcgctattg taaaattcat gttatatgga gggggcaaag
     1021 ttttcagggt gttgtttaga atgggaagat gtcccttgta tcaccatgga ccctcatgat
     1081 aattttgttt ctttcacttt ctactctgtt gacaaccatt gtctcctctt attttctttt
     1141 cattttctgt aactttttcg ttaaacttta gcttgcattt gtaacgaatt tttaaattca
     1201 cttttgttta tttgtcagat tgtaagtact ttctctaatc actttttttt caaggcaatc
     1261 agggtatatt atattgtact tcagcacagt tttagagaac aattgttata attaaatgat
     1321 aaggtagaat atttctgcat ataaattctg gctggcgtgg aaatattctt attggtagaa
     1381 acaactacat cctggtcatc atcctgcctt tctctttatg gttacaatga tatacactgt
     1441 ttgagatgag gataaaatac tctgagtcca aaccgggccc ctctgctaac catgttcatg
     1501 ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt
     1561 ttggcaaaga attgtaatac gactcactat agggcgaatt gatatgtcta gattagataa
     1621 aagtaaagtg attaacagcg cattagagct gcttaatgag gtcggaatcg aaggtttaac
     1681 aacccgtaaa ctcgcccaga agctaggtgt agagcagcct acattgtatt ggcatgtaaa
     1741 aaataagcgg gctttgctcg acgccttagc cattgagatg ttagataggc accatactca
     1801 cttttgccct ttagaagggg aaagctggca agatttttta cgtaataacg ctaaaagttt
     1861 tagatgtgct ttactaagtc atcgcgatgg agcaaaagta catttaggta cacggcctac
     1921 agaaaaacag tatgaaactc tcgaaaatca attagccttt ttatgccaac aaggtttttc
     1981 actagagaat gcattatatg cactcagcgc tgtggggcat tttactttag gttgcgtatt
     2041 ggaagatcaa gagcatcaag tcgctaaaga agaaagggaa acacctacta ctgatagtat
     2101 gccgccatta ttacgacaag ctatcgaatt atttgatcac caaggtgcag agccagcctt
     2161 cttattcggc cttgaattga tcatatgcgg attagaaaaa caacttaaat gtgaaagtgg
     2221 gtccggaccg ggtatggcgg cggcggttcg gatgaacatc cagatgctgc tggaggcggc
     2281 cgactatctg gagcggcggg agagagaagc tgaacatggt tatgcctcca tgttaccata
     2341 caataacaag gactaaggat cccgggaatt aattccgccc ctctccctcc ccccccccta
     2401 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt
     2461 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
     2521 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
     2581 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt
     2641 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
     2701 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
     2761 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     2821 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt
     2881 cgaggttaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac
     2941 acgatgataa tatgnnnggg gagcttggat atccattttc ggatctgatc aagagacagg
     3001 atgaggatcg tttcgcnnat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3061 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3121 gccgtgttgc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3181 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     3241 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3301 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     3361 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     3421 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     3481 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     3541 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     3601 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     3661 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     3721 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     3781 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     3841 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     3901 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     3961 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     4021 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4081 gtttgtccaa actcatcaat gtatcttatc atgtctggat cgatccgaag gtaagcctat
     4141 ccctaaccct ctcctcggtc tcgattctac gcgttgacat tgattattga ctagttatta
     4201 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     4261 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     4321 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     4381 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     4441 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     4501 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     4561 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     4621 tctccacccc attgacgtca atgggagttt gttttggaac caaaatcaac gggactttcc
     4681 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     4741 ggtctatata agcagagctc tccctatcag tgatagagat ctccctatca gtgatagaga
     4801 tctccctatc agtgatagag atctccctat cagtgataga gatcgagctg tttagtgaac
     4861 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgaatt
     4921 caggcctgtc gaccggtccg cggttaactc gagcggccgc taaacccgct gatcagcctc
     4981 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     5041 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     5101 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     5161 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     5221 aagaaccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5281 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5341 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5401 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5461 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5521 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5581 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5641 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5701 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5761 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5821 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5881 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5941 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6001 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6061 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6121 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6181 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6241 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6301 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6361 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6421 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6481 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6541 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6601 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6661 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6721 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6781 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6841 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6901 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6961 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7021 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7081 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7141 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7201 aaagtgccac ctgacgtc
//
