LOCUS Exported 7389 bp ds-DNA circular SYN 20-SEP-2017
DEFINITION synthetic circular DNA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7389)
TITLE Direct Submission
JOURNAL Exported Sep 20, 2017 from MiaoLingPlasmid
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..7389
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 131..334
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 375..398
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
misc_feature 472..589
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 638..1148
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
CDS 1170..1769
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
LTR 1897..2130
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 2202..2323
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin 2363..2498
/label=SV40 ori
/note="SV40 origin of replication"
rep_origin 2705..3160
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 3186..3290
/gene="bla"
/label=AmpR promoter
CDS 3291..4151
/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"
rep_origin 4322..4910
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 5379..5605
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 5606..5786
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 5833..5958
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 6451..6684
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
ORIGIN
1 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
61 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
121 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
181 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
241 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
301 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
361 gcgctaccgg tatggactac aaggatgacg atgacaaggg tggtggtggt ggtgcgtcga
421 cgatatcttc gaattctcga cctcgagaca aatggcagta ttcatccaca attttaaaag
481 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
541 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta
601 cagggacagc agagatccac tttggccgcg gctcgagggg gttggggttg cgccttttcc
661 aaggcagccc tgggtttgcg cagggacgcg gctgctctgg gcgtggttcc gggaaacgca
721 gcggcgccga ccctgggtct cgcacattct tcacgtccgt tcgcagcgtc acccggatct
781 tcgccgctac ccttgtgggc cccccggcga cgcttcctgc tccgccccta agtcgggaag
841 gttccttgcg gttcgcggcg tgccggacgt gacaaacgga agccgcacgt ctcactagta
901 ccctcgcaga cggacagcgc cagggagcaa tggcagcgcg ccgaccgcga tgggctgtgg
961 ccaatagcgg ctgctcagca gggcgcgccg agagcagcgg ccgggaaggg gcggtgcggg
1021 aggcggggtg tggggcggta gtgtgggccc tgttcctgcc cgcgcggtgt tccgcattct
1081 gcaagcctcc ggagcgcacg tcggcagtcg gctccctcgt tgaccgaatc accgacctct
1141 ctccccaggg ggatccaccg gagcttacca tgaccgagta caagcccacg gtgcgcctcg
1201 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
1261 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
1321 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
1381 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
1441 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
1501 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
1561 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
1621 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
1681 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
1741 ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg
1801 aaaggagcgc acgaccccat gcatcggtac ctttaagacc aatgacttac aaggcagctg
1861 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
1921 gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
1981 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
2041 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
2101 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
2161 atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt
2221 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
2281 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc
2341 tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
2401 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
2461 gctattccag aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc
2521 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa
2581 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
2641 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
2701 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
2761 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
2821 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
2881 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
2941 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
3001 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
3061 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
3121 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
3181 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
3241 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
3301 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
3361 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
3421 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
3481 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
3541 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
3601 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
3661 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
3721 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
3781 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
3841 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
3901 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
3961 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
4021 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
4081 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
4141 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
4201 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
4261 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
4321 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
4381 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
4441 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
4501 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
4561 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
4621 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
4681 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
4741 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
4801 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
4861 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
4921 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
4981 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
5041 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
5101 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
5161 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
5221 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
5281 gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc
5341 ctcactaaag ggaacaaaag ctggagctgc aagcttaatg tagtcttatg caatactctt
5401 gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc ttacaaggag agaaaaagca
5461 ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt gccttattag gaaggcaaca
5521 gacgggtctg acatggattg gacgaaccac tgaattgccg cattgcagag atattgtatt
5581 taagtgccta gctcgataca taaacgggtc tctctggtta gaccagatct gagcctggga
5641 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
5701 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
5761 ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg
5821 aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc
5881 gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga
5941 gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt
6001 cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg
6061 gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa
6121 atactgggac agctacaacc atcccttcag acaggatcag aagaacttag atcattatat
6181 aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa
6241 gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc
6301 gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg aattatataa
6361 atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt
6421 ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc
6481 agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt
6541 gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct
6601 gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag
6661 atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac
6721 cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca
6781 cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt
6841 aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa
6901 atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt
6961 cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt
7021 gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag
7081 gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc
7141 cattcgatta gtgaacggat ctcgacggta tcgatcacga gactagcctc gagcggccgc
7201 ccccttcacc gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc
7261 tgttagagag ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac
7321 gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat
7381 ggactatca
//