LOCUS Exported File 7139 bp ds-DNA circular SYN 28-1-2015
DEFINITION .
ACCESSION .
VERSION .
KEYWORDS pTet-On-Advanced
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7139)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2015-1-28 from SnapGene 2.0.1
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..7139
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 89..468
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 469..672
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 775..1521
/codon_start=1
/product="improved tetracycline-controlled transactivator"
/note="rtTA-Advanced"
/note="In the Tet-On(R) system, rtTA-Advanced binds to the
Tet-responsive element and stimulates transcription only in
the presence of tetracycline or doxycycline."
/translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
DALDDFDLDMLPADALDDFDLDMLPG"
polyA_signal 1531..1665
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(2385..2973)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3144..4004)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(4005..4109)
/gene="bla"
/note="AmpR promoter"
polyA_signal 4222..4356
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
CDS complement(4781..4801)
/codon_start=1
/locus_tag="
"
/product="nuclear localization signal of SV40 large T
antigen"
/note="SV40 NLS"
/note="
"
/protein_id="
"
/translation="PKKKRKV"
intron 4931..4996
/note="small t intron"
/note="simian virus 40 (SV40) small t antigen intron"
CDS complement(5416..6210)
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/note="NeoR/KanR"
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
promoter complement(6571..6900)
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 6585..6720
/note="SV40 ori"
/note="SV40 origin of replication"
ORIGIN
1 ctcgaggagc ttggcccatt gcatacgttg tatccatatc ataatatgta catttatatt
61 ggctcatgtc caacattacc gccatgttga cattgattat tgactagtta ttaatagtaa
121 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
181 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
241 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
301 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
361 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
421 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
481 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
541 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
601 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
661 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
721 gacctccata gaagacaccg ggaccgatcc agcctccgcg gccccgaatt caccatgtct
781 agactggaca agagcaaagt cataaacggc gctctggaat tactcaatgg agtcggtatc
841 gaaggcctga cgacaaggaa actcgctcaa aagctgggag ttgagcagcc taccctgtac
901 tggcacgtga agaacaagcg ggccctgctc gatgccctgc caatcgagat gctggacagg
961 catcataccc acttctgccc cctggaaggc gagtcatggc aagactttct gcggaacaac
1021 gccaagtcat tccgctgtgc tctcctctca catcgcgacg gggctaaagt gcatctcggc
1081 acccgcccaa cagagaaaca gtacgaaacc ctggaaaatc agctcgcgtt cctgtgtcag
1141 caaggcttct ccctggagaa cgcactgtac gctctgtccg ccgtgggcca ctttacactg
1201 ggctgcgtat tggaggaaca ggagcatcaa gtagcaaaag aggaaagaga gacacctacc
1261 accgattcta tgcccccact tctgagacaa gcaattgagc tgttcgaccg gcagggagcc
1321 gaacctgcct tccttttcgg cctggaacta atcatatgtg gcctggagaa acagctaaag
1381 tgcgaaagcg gcgggccggc cgacgccctt gacgattttg acttagacat gctcccagcc
1441 gatgcccttg acgactttga ccttgatatg ctgcctgctg acgctcttga cgattttgac
1501 cttgacatgc tccccgggta actaagtaag gatccagaca tgataagata cattgatgag
1561 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
1621 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
1681 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
1741 ctctacaaat gtggtatggc tgattatgat cctgcaagcc tcgtcgtcct ggccggacca
1801 cgctatctgt gcaaggtccc cggccccgga cgcgcgctcc atgagcagag cgcccgccgc
1861 cgaggcgaag actcgggcgg cgccctgccc gtcccaccag gtcaacaggc ggtaaccggc
1921 ctcttcatcg ggaatgcgcg cgaccttcag catcgccggc atgtccccct ggcggacggg
1981 aagtatccag ctcgaccaag cttggcgaga ttttcaggag ctaaggaagc taaaatggag
2041 aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
2101 gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct gcattaatga
2161 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
2221 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
2281 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
2341 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
2401 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
2461 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
2521 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
2581 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
2641 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
2701 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
2761 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
2821 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
2881 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
2941 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
3001 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
3061 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
3121 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
3181 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
3241 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
3301 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
3361 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
3421 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
3481 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
3541 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
3601 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
3661 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
3721 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca
3781 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
3841 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
3901 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
3961 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
4021 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
4081 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
4141 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
4201 cgtcttcact cgaggtcgag ggatccagac atgataagat acattgatga gtttggacaa
4261 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
4321 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
4381 atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa
4441 tgtggtatgg ctgattatga tctctagtca aggcactata catcaaatat tccttattaa
4501 cccctttaca aattaaaaag ctaaaggtac acaatttttg agcatagtta ttaatagcag
4561 acactctatg cctgtgtgga gtaagaaaaa acagtatgtt atgattataa ctgttatgcc
4621 tacttataaa ggttacagaa tatttttcca taattttctt gtatagcagt gcagcttttt
4681 cctttgtggt gtaaatagca aagcaagcaa gagttctatt actaaacaca gcatgactca
4741 aaaaacttag caattctgaa ggaaagtcct tggggtcttc tacctttctc ttcttttttg
4801 gaggagtaga atgttgagag tcagcagtag cctcatcatc actagatggc atttcttctg
4861 agcaaaacag gttttcctca ttaaaggcat tccaccactg ctcccattca tcagttccat
4921 aggttggaat ctaaaataca caaacaatta gaatcagtag tttaacacat tatacactta
4981 aaaattttat atttacctta gagctttaaa tctctgtagg tagtttgtcc aattatgtca
5041 caccacagaa gtaaggttcc ttcacaaaga tccggaccaa agcggccatc gtgcctcccc
5101 actcctgcag ttcgggggca tggatgcgcg gatagccgct gctggtttcc tggatgccga
5161 cggatttgca ctgccggtag aactccgcga ggtcgtccag cctcaggcag cagctgaacc
5221 aactcgcgag gggatcgagc ccggggtggg cgaagaactc cagcatgaga tccccgcgct
5281 ggaggatcat ccagccggcg tcccggaaaa cgattccgaa gcccaacctt tcatagaagg
5341 cggcggtgga atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga
5401 accccagagt cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga
5461 atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc
5521 ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg
5581 gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc
5641 atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa
5701 cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc
5761 ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca
5821 ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc
5881 ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca
5941 gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc
6001 cagccacgat agccgcgctg cctcgtcctg cagttcattc agggcaccgg acaggtcggt
6061 cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca
6121 gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga
6181 acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc
6241 agatcttgat cccctgcgcc atcagatcct tggcggcaag aaagccatcc agtttacttt
6301 gcagggcttc ccaaccttac cagagggcgc cccagctggc aattccggtt cgcttgctgt
6361 ccataaaacc gcccagtcta gctatcgcca tgtaagccca ctgcaagcta cctgctttct
6421 ctttgcgctt gcgttttccc ttgtccagat agcccagtag ctgacattca tccggggtca
6481 gcaccgtttc tgcggactgg ctttctacgt gttccgcttc ctttagcagc ccttgcgccc
6541 tgagtgcttg cggcagcgtg aagctagctt tttgcaaaag cctaggcctc caaaaaagcc
6601 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
6661 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
6721 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact
6781 tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat
6841 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac
6901 attccacagc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
6961 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
7021 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
7081 cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcacct
//