LOCUS       40924_16425       10049 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pe2TetOn(PacI), complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10049)
  AUTHORS   Backman CM, Zhang Y, Hoffer BJ, Tomac AC.
  TITLE     Tetracycline-inducible expression systems for the generation of 
            transgenic animals: a comparison of various inducible systems 
            carried in a single vector
  JOURNAL   J. Neurosci. Methods 139 (2), 257-262 (2004)
  PUBMED    15488239
REFERENCE   2  (bases 1 to 10049)
  AUTHORS   Backman CM, Zhang Y, Hoffer BJ, Tomac AC.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-DEC-2005) DHHS, NIDA/NIH, 5500 Nathan Shock Drive, 
            Baltimore, MD 21224, USA
REFERENCE   3  (bases 1 to 10049)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10049)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. 
            Neurosci. Methods"; date: "2004"; volume: "139"; issue: "2"; pages: 
            "257-262"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (19-DEC-2005) DHHS, NIDA/NIH, 5500 Nathan Shock Drive, Baltimore, MD
            21224, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10049
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(78..458)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    714..1300
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     1329..1345
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1368..2111
                     /codon_start=1
                     /label=rtTA-Advanced
                     /note="improved tetracycline-controlled transactivator"
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     polyA_signal    complement(2124..2258)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    6799..6933
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    6963..7430
                     /label=contains SV40 polyA signal
                     /note="contains SV40 polyA signal"
     polyA_signal    7289..7423
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(7507..7821)
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     promoter        complement(7861..7879)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7900..7916)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7924..7940
                     /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(7948..7978)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7993..8014)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(8302..8890)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(9064..9921)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(9922..10026)
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
      661 gccccccctc gagaattcac gcgtcgagca tgcatctagg gcggccaatt ccgcccctct
      721 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
      781 gtctatatgt gattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
      841 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
      901 ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg
      961 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
     1021 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
     1081 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg
     1141 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc
     1201 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg
     1261 gttttccttt gaaaaacacg atgataagct tgccacaacc cgggatcctc tagagtcgac
     1321 cgggccgctc tagaactagt ggatcccccg ggctgcagga attcaccatg tctagactgg
     1381 acaagagcaa agtcataaac ggcgctctgg aattactcaa tggagtcggt atcgaaggcc
     1441 tgacgacaag gaaactcgct caaaagctgg gagttgagca gcctaccctg tactggcacg
     1501 tgaagaacaa gcgggccctg ctcgatgccc tgccaatcga gatgctggac aggcatcata
     1561 cccacttctg ccccctggaa ggcgagtcat ggcaagactt tctgcggaac aacgccaagt
     1621 cattccgctg tgctctcctc tcacatcgcg acggggctaa agtgcatctc ggcacccgcc
     1681 caacagagaa acagtacgaa accctggaaa atcagctcgc gttcctgtgt cagcaaggct
     1741 tctccctgga gaacgcactg tacgctctgt ccgccgtggg ccactttaca ctgggctgcg
     1801 tattggagga acaggagcat caagtagcaa aagaggaaag agagacacct accaccgatt
     1861 ctatgccccc acttctgaga caagcaattg agctgttcga ccggcaggga gccgaacctg
     1921 ccttcctttt cggcctggaa ctaatcatat gtggcctgga gaaacagcta aagtgcgaaa
     1981 gcggcgggcc ggccgacgcc cttgacgatt ttgacttaga catgctccca gccgatgccc
     2041 ttgacgactt tgaccttgat atgctgcctg ctgacgctct tgacgatttt gaccttgaca
     2101 tgctccccgg gtaactaagt aaggatccag acatgataag atacattgat gagtttggac
     2161 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     2221 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     2281 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     2341 aatgtggtat ggctgattat gatcctgcaa gcctcgtcgt ctggccggac cacgctatct
     2401 gtgcaaggtc cccggacgcg cgctccatga gcagagcgcc cgccgccgag gcaagactcg
     2461 ggcggcgccc tgcccgtccc accaggtcaa caggcggtaa ccggcctctt catcgggaat
     2521 gcgcgcgacc ttcagcatcg ccggcatgtc ccctggcgga cgggaagtat cagctcgacc
     2581 aagctcggtc gagacaccct gctttggtct ctctaaagct ggagcgacag ctgctcatca
     2641 ccatgtttga cttaagcagc tttcagtgta gactctcggc cattgttttg agtccattta
     2701 ttgatactga ctagtctccc aatcctgtca cctctttcaa agatacaatc cccataggat
     2761 gctgggcagg tcttatctac tcacggagat ctgtttgctc ttggaacttt tgtgttctaa
     2821 tcattgcacc ctatttatat attttagtaa ctgaaagtca gttcttcccc ttcactggcc
     2881 tactgactct tgaatgtaag tgatatctta tttactatat ttctttcagc tctttgataa
     2941 cgtaatggga tttttggtca tatgggaatc accgaatttt aaggaaagga gggtgcttcc
     3001 ccttccagag ccttcagtct ctctttctgt ggagcacaag ggttttatca aagcagtgct
     3061 gtacttgggg tggttaataa aggaaggaat taaatgcccc cacctccctc ccctctgcca
     3121 ccccttcacc catgcaccct cacctgactc ctagcctttc atctgaaggc aggcagttgc
     3181 tcaggacctg accttgtgca tgtctcgtga cctcatccta tgacctcatc ctatgacctc
     3241 atcctggggt tttttcattt taacctctag ctcacaccgg cgtattctgc tctgtggtgt
     3301 gcagagggca gagctgagct aagagattca gctatatttt ctttctattc ccattcagag
     3361 atggtacaag cggtagagtc aaaagttcac tctgtgtgtg tgtgtgtgtg tgtgtgtgtg
     3421 tgtgtgtatc tccctcaggt atcttccctc agtaccaccc aacttttttt tgaaagggcc
     3481 gctcactggc ctagataggc taggctgact gaccactgag ctggggaacc cacttgtccc
     3541 catctcctca aactgggatt gcaagcttag gccaccacac ctggatttct attcttggat
     3601 tctgtgatct gaactcaggt cctcatgttt gcaaggcaaa tcttttctca actgagctat
     3661 tattctagcc ctggaacttc attcttctat ggggttgact ccccaagtct tacaggaaat
     3721 ataccacacc ttgagagtag ttcccagtga ggaagcaatt tgggcccatc ctgagagaca
     3781 ggaatcctac aggcattccc ctgagggggc cactggacta taggaacgtg tttggaatca
     3841 gtgacaatcc agaaggtcac acagctcagt ctagaggagc ccttgcctgt acttgttcta
     3901 attctacatc agtttataaa tagcatccgg ccgactgccc agtgatgact tgggtcatcc
     3961 ctcctcaggg tccttgtagc tggggtgtga tttttctgtg ggtccccttg acctcagaag
     4021 cagaggacca tggaggacaa caggattgct gctggtgctg tcagcagctg acatctcgtc
     4081 acaccctgcc caccactgtt ctccgcactc tctgcagtta gttttcataa gccttatgag
     4141 gcagctgctc ttctgattct cacattacaa gtgaggcaga cagtcccagg aatgcttagg
     4201 agagatgatg cagaaatgca cagtctgctg tgtatccctt gagctgctcc ttggtgtaac
     4261 tgtatggaag ttgaaactca agcacctgtg tagtctctca gctctaccca caggagcttt
     4321 gcaacatacg gtcccccaaa cctgaatctt cccctatgga atggagccat ggtacttaat
     4381 ttatagttta tgaggaaaag atacaggcta aaagttcaga gctctacctg gcacaggaga
     4441 ttctcagagg gcaaccagtt ctctctactt gcatggcatt ttggagcctg agtttctaat
     4501 atgtatgtga agccattact ctgcatcttg gattgctgtg tgctctctga gcttggttca
     4561 agtgttcagc atggctgtgt ttccaccttt gccactgtgt ggctctgggt gggcatggct
     4621 aagcaccctc ctgcctgaag caaggtcttc tgggtttcat gaggactcag gaggagcctt
     4681 gtttcattcc tggaatggtg gtcctccctt ctctatggca tagctcttct catagtgtgg
     4741 tgctcccctg tgttatcttc ttacactgtg gtactcctcc actgttgtgt gctgcacaga
     4801 gctagtgctc ccctgaaagt gtgtctgtct ctttgtgggt attgcaaaaa cagagctgtt
     4861 tttctttctg tactgaggta cctgttattc ctccaggaga gagaggccag acaggccaca
     4921 agaggatggt ccttgtagtt tctagtggtt agaatgagga tagccaccat aggctcctgt
     4981 atttgatgtc cagtccttag ttagcaaaac tgtttgggaa ggattagaag gtgtggcctt
     5041 gttggaggag gtgtgtattg gaggtgggct tgggtttcca aaagcctaca ctaggctttt
     5101 tctctgcctg caaatcagac tgtaaagttc ttagctactg cttgagcgtg cctgcttccc
     5161 accatgatgt acgagcatgt agtaaccctc taaaactaag caagcacata attaaatgct
     5221 ttcttcttta agagttgcct tggtcatagt gtctcttcac agcaatagag tagaaactga
     5281 gacacaggta ttgggcaagg tctttgagaa agggagggtg ctagggaatc gggcaggcat
     5341 ctaggtaggc ctgtaaggca tccagaaagc ccggaggaga aaagtctttg gtgcagctgg
     5401 ataagaagct ggtgaagagg gaaatggttt taactttagt aggtgagcct gacacttgct
     5461 cttcagacct cttctgcaga gtacagcagc agtagggcat tgcctttttg tcaggtaata
     5521 tcacatagta ggacttacaa tagggctcac agtagggctt acaatagggc tcatagaaat
     5581 ggcagttctt gtcataaatt tagttgctga tgattaaaac accatcagcc tgaagacaga
     5641 aaagactgga caagaggatc tgctcttggt ccaaggaagt agtgcaggtc ctgaaggatg
     5701 ctgggaacat gcagtggtgg ccttatgtgt gccagatgtg gctgagagca gctgttagct
     5761 gcacttccac ggtgcttgaa tgtgaccccc cccagccccc actcttctgt gttttaaata
     5821 tttgttctgt ggctggtagc actatctgga gaagctgggg gcctctatag gggtaaagtc
     5881 tagctagaag aaacagacat ttgcaggtta tacctagcac caggttacag ccactctcgg
     5941 ctacctgtag acagcttctt cacatgttct caccactaag aatgctgcca tgcctccccc
     6001 ccccaccccc taccccccac ccccaatgat ggactaaaac atcaagccct aatacaattt
     6061 cctcctttga cttgcttctg tgaggtattt tgttcacaga ggtgaaaaaa aaaaactaac
     6121 agaagattgg tgccaaagaa gcaattgggt cattgctgtg actatgctga catccatggg
     6181 tccttagaac tggcttgcag agggactggg gagagtttag agatgtagga tctgggcagt
     6241 gggtggttct ggtatgagta aggaaaacca caatggttca gatggaagct ggaactgctg
     6301 ataagttgac tagggaccat ttgtgttaca tacatttgtg aatatacatt cagccatgtt
     6361 aggcttgtat ccaggacatt tgaatgaggc tggactgaca agtatggcat aatggtgggg
     6421 aaatttcaag gcagcataga attcggtcga cccgggcggc cagcttggtc gagctggata
     6481 cttcccgtcc gccaggggga catgccggcg atgctgaagg tcgcgcgcat tcccgatgaa
     6541 gaggccggtt accgcctgtt gacctggtgg gacgggcagg gcgccgcccg agtcttcgcc
     6601 tcggcggcgg gcgctctgct catggagcgc gcgtccgggg ccggggacct tgcacagata
     6661 gcgtggtccg gccaggacga cgaggcttgc aggatcataa tcagccatac cacatttgta
     6721 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     6781 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     6841 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     6901 aaactcatca atgtatctta tcatgtctgg atcctctaga ggatccccgg gtaccgagct
     6961 cagcttggtc gagctgatac ttcccgtccg ccaggggaca tgccggcgat gctgaaggtc
     7021 gcgcgcattc ccgatgaaga ggccggttac cgcctgttga cctggtggga cgggcagggc
     7081 gccgcccgag tcttgcctcg gcggcgggcg ctctgctcat ggagcgcgcg tccggggacc
     7141 ttgcacagat agcgtggtcc ggccagacga cgaggcttgc aggatcataa tcagccatac
     7201 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     7261 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     7321 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     7381 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcctctaga attgttaatt
     7441 aacgttaatt aacgttaatt aacgttaatt aacgttaatt aacgggtacc gagctcgaat
     7501 tctccaggcg atctgacggt tcactaaacg agctctgctt atataggcct cccaccgtac
     7561 acgcctacct cgacatacgt tctctatcac tgatagggag taaactcgac atacgttctc
     7621 tatcactgat agggataaac tcgacatacg ttctctatca ctgataggga gtaaactcga
     7681 catacgttct ctatcactga tagggagtaa actcgacata cgttctctat cactgatagg
     7741 gagtaaactc gacatcgttc tctatcactg atagggagta aactcgacat acgttctcta
     7801 tcactgatag ggagtaaact cgaagcggcc gccaccgcgg tggagctcca gcttttgttc
     7861 cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     7921 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     7981 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     8041 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     8101 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     8161 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     8221 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     8281 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     8341 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     8401 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     8461 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     8521 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     8581 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     8641 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     8701 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     8761 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     8821 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     8881 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     8941 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     9001 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     9061 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     9121 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     9181 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     9241 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     9301 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     9361 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     9421 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     9481 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     9541 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     9601 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     9661 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     9721 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     9781 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     9841 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     9901 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     9961 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
    10021 tccgcgcaca tttccccgaa aagtgccac
//
