LOCUS Exported 7790 bp ds-DNA circular SYN 27-AUG-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 26
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7790)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Saturday, August 27, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..7790
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 1..634
/note="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/note="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1303..1536
/note="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 2028..2143
/note="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)"
promoter 2206..2520
/note="tight TRE promoter"
/note="Tet-responsive promoter PTight, consisting of seven
tet operator sequences followed by the minimal CMV
promoter"
protein_bind 2214..2232
/gene="tetO"
/bound_moiety="tetracycline repressor TetR"
/note="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2250..2268
/gene="tetO"
/bound_moiety="tetracycline repressor TetR"
/note="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2285..2303
/gene="tetO"
/bound_moiety="tetracycline repressor TetR"
/note="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2321..2339
/gene="tetO"
/bound_moiety="tetracycline repressor TetR"
/note="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2357..2375
/gene="tetO"
/bound_moiety="tetracycline repressor TetR"
/note="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2392..2410
/gene="tetO"
/bound_moiety="tetracycline repressor TetR"
/note="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2428..2446
/gene="tetO"
/bound_moiety="tetracycline repressor TetR"
/note="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
promoter 2576..3075
/note="PGK promoter"
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 3096..3695
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/note="PuroR"
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
misc_feature 3712..4300
/note="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS complement(4183..4194)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/note="Factor Xa site"
/translation="IEGR"
LTR 4507..5140
/note="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
primer_bind complement(5268..5284)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 5292..5308
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/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(5316..5346)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 5361..5382
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5670..6258)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6429..7289)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(7290..7394)
/gene="bla"
/note="AmpR promoter"
polyA_signal 7442..7576
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
ORIGIN
1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
2161 acagcagaga tccagtttat cgatgaggcc ctttcgtctt cactcgagtt tactccctat
2221 cagtgataga gaacgtatgt cgagtttact ccctatcagt gatagagaac gatgtcgagt
2281 ttactcccta tcagtgatag agaacgtatg tcgagtttac tccctatcag tgatagagaa
2341 cgtatgtcga gtttactccc tatcagtgat agagaacgta tgtcgagttt atccctatca
2401 gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacgt atgtcgaggt
2461 aggcgtgtac ggtgggaggc ctatataagc agagctcgtt tagtgaaccg tcagatcgcc
2521 tggagaagga tccgcggccg cgccggctct agatcgcgaa cgcgtgaatt ctaccgggta
2581 ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc gctgggcact
2641 tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg taggcgccaa
2701 ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct agtcaggaag
2761 ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc
2821 actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg cctttggggc
2881 agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg aaggggtggg
2941 tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga
3001 ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca
3061 tctccgggcc tttcgacctg cagcccaagc ttaccatgac cgagtacaag cccacggtgc
3121 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
3181 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
3241 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
3301 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
3361 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
3421 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
3481 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
3541 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
3601 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
3661 gcacctggtg catgacccgc aagcccggtg cctgacgggc gcgtctggaa caatcaacct
3721 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
3781 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
3841 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
3901 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
3961 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
4021 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
4081 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
4141 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
4201 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
4261 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattaa ttctgcagtc
4321 gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc aatgctgatt
4381 gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca cctcaggtac
4441 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaaga
4501 ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat ctgtggatct
4561 accacacaca aggctacttc cctgattagc agaactacac accagggcca ggggtcagat
4621 atccactgac ctttggatgg tgctacaagc tagtaccagt tgagccagat aaggtagaag
4681 aggccaataa aggagagaac accagcttgt tacaccctgt gagcctgcat gggatggatg
4741 acccggagag agaagtgtta gagtggaggt ttgacagccg cctagcattt catcacgtgg
4801 cccgagagct gcatccggag tacttcaaga actgctgata tcgagcttgc tacaagggac
4861 tttccgctgg ggactttcca gggaggcgtg gcctgggcgg gactggggag tggcgagccc
4921 tcagatcctg catataagca gctgcttttt gcctgtactg ggtctctctg gttagaccag
4981 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
5041 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
5101 tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt
5161 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gccttgacat
5221 tgctagcgtt taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
5281 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
5341 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
5401 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
5461 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
5521 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
5581 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
5641 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
5701 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
5761 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
5821 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
5881 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
5941 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
6001 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
6061 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
6121 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
6181 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
6241 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
6301 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
6361 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
6421 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
6481 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
6541 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
6601 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
6661 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
6721 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
6781 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
6841 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
6901 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
6961 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
7021 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
7081 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
7141 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
7201 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
7261 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
7321 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
7381 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
7441 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
7501 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
7561 ttatcatgtc tggatcaact ggataactca agctaaccaa aatcatccca aacttcccac
7621 cccataccct attaccactg ccaattacct gtggtttcat ttactctaaa cctgtgattc
7681 ctctgaatta ttttcatttt aaagaaattg tatttgttaa atatgtacta caaacttagt
7741 agtttttaaa gaaattgtat ttgttaaata tgtactacaa acttagtagt
//