LOCUS Exported 9100 bp ds-DNA circular SYN 27-AUG-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 35
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9100)
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..9100
/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..2555
/note="TRE3GV promoter"
/note="3rd-generation Tet-responsive promoter that can be
activated by binding of Tet-On(R) 3G, optimized for
retroviral and lentiviral vectors"
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 2286..2304
/gene="tetO"
/bound_moiety="tetracycline repressor TetR"
/note="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2322..2340
/gene="tetO"
/bound_moiety="tetracycline repressor TetR"
/note="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2358..2376
/gene="tetO"
/bound_moiety="tetracycline repressor TetR"
/note="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2394..2412
/gene="tetO"
/bound_moiety="tetracycline repressor TetR"
/note="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2430..2448
/gene="tetO"
/bound_moiety="tetracycline repressor TetR"
/note="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
misc_feature 3275..3849
/note="IRES"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
misc_feature 3299..3851
/note="IRES"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
promoter 3885..4384
/note="PGK promoter"
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 4405..5004
/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 5022..5610
/note="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS complement(5493..5504)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/note="Factor Xa site"
/translation="IEGR"
LTR 5818..6451
/note="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
primer_bind complement(6580..6596)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 6604..6620
/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(6628..6658)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 6673..6694
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(6982..7567)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7738..8598)
/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(8599..8703)
/gene="bla"
/note="AmpR promoter"
polyA_signal 8751..8885
/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 gaacgtatga agagtttact ccctatcagt gatagagaac gtatgcagac
2281 tttactccct atcagtgata gagaacgtat aaggagttta ctccctatca gtgatagaga
2341 acgtatgacc agtttactcc ctatcagtga tagagaacgt atctacagtt tactccctat
2401 cagtgataga gaacgtatat ccagtttact ccctatcagt gatagagaac gtatgtcgag
2461 gtaggcgtgt acggtgggcg cctataaaag cagagctcgt ttagtgaacc gtcagatcgc
2521 ctggagcaat tccacaacac ttttgtctta tacttggatc accatggctc agtcaaagca
2581 cggcctgacc aaggagatga ccatgaagta ccgcatggag ggctgcgtgg acggccacaa
2641 gttcgtgatc accggcgagg gcatcggcta ccccttcaag ggcaagcagg ccatcaacct
2701 gtgcgtggtg gagggcggcc ccttgccctt cgccgaggac atcttgtccg ccgccttcat
2761 gtacggcaac cgcgtgttca ccgagtaccc ccaggacatc gtcgactact tcaagaactc
2821 ctgccccgcc ggctacacct gggaccgctc cttcctgttc gaggacggcg ccgtgtgcat
2881 ctgcaacgcc gacatcaccg tgagcgtgga ggagaactgc atgtaccacg agtccaagtt
2941 ctacggcgtg aacttccccg ccgacggccc cgtgatgaag aagatgaccg acaactggga
3001 gccctcctgc gagaagatca tccccgtgcc caagcagggc atcttgaagg gcgacgtgag
3061 catgtacctg ctgctgaagg acggtggccg cttgcgctgc cagttcgaca ccgtgtacaa
3121 ggccaagtcc gtgccccgca agatgcccga ctggcacttc atccagcaca agctgacccg
3181 cgaggaccgc agcgacgcca agaaccagaa gtggcacctg accgagcacg ccatcgcctc
3241 cggatctgca ttgccctgag gccgccccgg ccggcccctc tccctccccc cccccctaac
3301 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
3361 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
3421 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
3481 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
3541 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
3601 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
3661 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
3721 ccccattgta tgggatctga tctggggcct cggtacacat gctttacatg tgtttagtcg
3781 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
3841 cgatgataat atggcccccg ggacgcgtca tatggaattc taccgggtag gggaggcgct
3901 tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac
3961 aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt
4021 ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc
4081 cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt
4141 gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata
4201 gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg
4261 gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag gcccggcatt
4321 ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat ctccgggcct
4381 ttcgacctgc agcccaagct taccatgacc gagtacaagc ccacggtgcg cctcgccacc
4441 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
4501 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
4561 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
4621 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
4681 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
4741 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
4801 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
4861 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
4921 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
4981 atgacccgca agcccggtgc ctgacggggc gcgtctggaa caatcaacct ctggattaca
5041 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
5101 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
5161 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
5221 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
5281 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
5341 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
5401 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga
5461 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
5521 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
5581 gtcggatctc cctttgggcc gcctccccgc ctggaattaa ttctgcagtc gagacctaga
5641 aaaacatgga gcaatcacaa gtagcaatac agcagctacc aatgctgatt gtgcctggct
5701 agaagcacaa gaggaggagg aggtgggttt ttccagtcac acctcaggta cctttaagac
5761 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag aggggactgg
5821 aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac
5881 aaggctactt ccctgattag cagaactaca caccagggcc aggggtcaga tatccactga
5941 cctttggatg gtgctacaag ctagtaccag ttgagccaga taaggtagaa gaggccaata
6001 aaggagagaa caccagcttg ttacaccctg tgagcctgca tgggatggat gacccggaga
6061 gagaagtgtt agagtggagg tttgacagcc gcctagcatt tcatcacgtg gcccgagagc
6121 tgcatccgga gtacttcaag aactgctgat atcgagcttg ctacaaggga ctttccgctg
6181 gggactttcc agggaggcgt ggcctgggcg ggactgggga gtggcgagcc ctcagatcct
6241 gcatataagc agctgctttt tgcctgtact gggtctctct ggttagacca gatctgagcc
6301 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
6361 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
6421 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag
6481 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggccttgaca ttgctagcgt
6541 tttaccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
6601 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
6661 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
6721 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
6781 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
6841 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
6901 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
6961 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
7021 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
7081 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
7141 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
7201 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
7261 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
7321 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
7381 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
7441 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
7501 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
7561 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
7621 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
7681 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
7741 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
7801 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
7861 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
7921 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
7981 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
8041 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
8101 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
8161 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
8221 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
8281 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
8341 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
8401 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
8461 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
8521 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
8581 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
8641 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
8701 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc aacttgttta
8761 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
8821 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
8881 ggatcaactg gataactcaa gctaaccaaa atcatcccaa acttcccacc ccatacccta
8941 ttaccactgc caattaccta gtggtttcat ttactctaaa cctgtgattc ctctgaatta
9001 ttttcatttt aaagaaattg tatttgttaa atatgtacta caaacttagt agtttttaaa
9061 gaaattgtat ttgttaaata tgtactacaa acttagtagt
//