LOCUS Exported 6357 bp ds-DNA circular SYN 24-SEP-2017
DEFINITION synthetic circular DNA
KEYWORDS psiCHECK2-let-7 4x
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6357)
TITLE Direct Submission
JOURNAL Exported Sep 24, 2017 from MiaoLingPlasmid
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..6357
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 62..419
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 270..405
/label=SV40 ori
/note="SV40 origin of replication"
intron 489..621
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
promoter 666..684
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 694..1629
/codon_start=1
/product="Renilla luciferase"
/label=hRluc
/note="human codon-optimized"
/translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
misc_feature 1644..1751
/label=let-7 4x
polyA_signal 1772..1820
/note="synthetic polyadenylation signal"
promoter 1829..2580
/label=HSV TK promoter
/note="herpes simplex virus thymidine kinase promoter"
CDS 2616..4268
/codon_start=1
/label=hluc+
/translation="MADAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
polyA_signal 4312..4433
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter 4566..4670
/gene="bla"
/label=AmpR promoter
CDS 4671..5531
/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 5702..6290
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 agatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct
61 tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
241 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
301 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
361 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
421 agcttgattc ttctgacaca acagtctcga acttaagctg cagaagttgg tcgtgaggca
481 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
541 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
601 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga
661 gtacttaata cgactcacta taggctagcc accatggctt ccaaggtgta cgaccccgag
721 caacgcaaac gcatgatcac tgggcctcag tggtgggctc gctgcaagca aatgaacgtg
781 ctggactcct tcatcaacta ctatgattcc gagaagcacg ccgagaacgc cgtgattttt
841 ctgcatggta acgctgcctc cagctacctg tggaggcacg tcgtgcctca catcgagccc
901 gtggctagat gcatcatccc tgatctgatc ggaatgggta agtccggcaa gagcgggaat
961 ggctcatatc gcctcctgga tcactacaag tacctcaccg cttggttcga gctgctgaac
1021 cttccaaaga aaatcatctt tgtgggccac gactgggggg cttgtctggc ctttcactac
1081 tcctacgagc accaagacaa gatcaaggcc atcgtccatg ctgagagtgt cgtggacgtg
1141 atcgagtcct gggacgagtg gcctgacatc gaggaggata tcgccctgat caagagcgaa
1201 gagggcgaga aaatggtgct tgagaataac ttcttcgtcg agaccatgct cccaagcaag
1261 atcatgcgga aactggagcc tgaggagttc gctgcctacc tggagccatt caaggagaag
1321 ggcgaggtta gacggcctac cctctcctgg cctcgcgaga tccctctcgt taagggaggc
1381 aagcccgacg tcgtccagat tgtccgcaac tacaacgcct accttcgggc cagcgacgat
1441 ctgcctaaga tgttcatcga gtccgaccct gggttctttt ccaacgctat tgtcgaggga
1501 gctaagaagt tccctaacac cgagttcgtg aaggtgaagg gcctccactt cagccaggag
1561 gacgctccag atgaaatggg taagtacatc aagagcttcg tggagcgcgt gctgaagaac
1621 gagcagtaat tctaggcgat cgctcgagac tatacaagga tctacctcag tcgagactat
1681 acaaggatct acctcagtcg agactataca aggatctacc tcagtcgaga ctatacaagg
1741 atctacctca gtcgacgcgg ccgctggccg caataaaata tctttatttt cattacatct
1801 gtgtgttggt tttttgtgtg aggatctaaa tgagtcttcg gacctcgcgg gggccgctta
1861 agcggtggtt agggtttgtc tgacgcgggg ggagggggaa ggaacgaaac actctcattc
1921 ggaggcggct cggggtttgg tcttggtggc cacgggcacg cagaagagcg ccgcgatcct
1981 cttaagcacc cccccgccct ccgtggaggc gggggtttgg tcggcgggtg gtaactggcg
2041 ggccgctgac tcgggcgggt cgcgcgcccc agagtgtgac cttttcggtc tgctcgcaga
2101 cccccgggcg gcgccgccgc ggcggcgacg ggctcgctgg gtcctaggct ccatggggac
2161 cgtatacgtg gacaggctct ggagcatccg cacgactgcg gtgatattac cggagacctt
2221 ctgcgggacg agccgggtca cgcggctgac gcggagcgtc cgttgggcga caaacaccag
2281 gacggggcac aggtacacta tcttgtcacc cggaggcgcg agggactgca ggagcttcag
2341 ggagtggcgc agctgcttca tccccgtggc ccgttgctcg cgtttgctgg cggtgtcccc
2401 ggaagaaata tatttgcatg tctttagttc tatgatgaca caaaccccgc ccagcgtctt
2461 gtcattggcg aattcgaaca cgcagatgca gtcggggcgg cgcggtccca ggtccacttc
2521 gcatattaag gtgacgcgtg tggcctcgaa caccgagcga ccctgcagcg acccgcttaa
2581 aagcttggca ttccggtact gttggtaaag ccaccatggc cgatgctaag aacattaaga
2641 agggccctgc tcccttctac cctctggagg atggcaccgc tggcgagcag ctgcacaagg
2701 ccatgaagag gtatgccctg gtgcctggca ccattgcctt caccgatgcc cacattgagg
2761 tggacatcac ctatgccgag tacttcgaga tgtctgtgcg cctggccgag gccatgaaga
2821 ggtacggcct gaacaccaac caccgcatcg tggtgtgctc tgagaactct ctgcagttct
2881 tcatgccagt gctgggcgcc ctgttcatcg gagtggccgt ggcccctgct aacgacattt
2941 acaacgagcg cgagctgctg aacagcatgg gcatttctca gcctaccgtg gtgttcgtgt
3001 ctaagaaggg cctgcagaag atcctgaacg tgcagaagaa gctgcctatc atccagaaga
3061 tcatcatcat ggactctaag accgactacc agggcttcca gagcatgtac acattcgtga
3121 catctcatct gcctcctggc ttcaacgagt acgacttcgt gccagagtct ttcgacaggg
3181 acaaaaccat tgccctgatc atgaacagct ctgggtctac cggcctgcct aagggcgtgg
3241 ccctgcctca tcgcaccgcc tgtgtgcgct tctctcacgc ccgcgaccct attttcggca
3301 accagatcat ccccgacacc gctattctga gcgtggtgcc attccaccac ggcttcggca
3361 tgttcaccac cctgggctac ctgatttgcg gctttcgggt ggtgctgatg taccgcttcg
3421 aggaggagct gttcctgcgc agcctgcaag actacaaaat tcagtctgcc ctgctggtgc
3481 caaccctgtt cagcttcttc gctaagagca ccctgatcga caagtacgac ctgtctaacc
3541 tgcacgagat tgcctctggc ggcgccccac tgtctaagga ggtgggcgaa gccgtggcca
3601 agcgctttca tctgccaggc atccgccagg gctacggcct gaccgagaca accagcgcca
3661 ttctgattac cccagagggc gacgacaagc ctggcgccgt gggcaaggtg gtgccattct
3721 tcgaggccaa ggtggtggac ctggacaccg gcaagaccct gggagtgaac cagcgcggcg
3781 agctgtgtgt gcgcggccct atgattatgt ccggctacgt gaataaccct gaggccacaa
3841 acgccctgat cgacaaggac ggctggctgc actctggcga cattgcctac tgggacgagg
3901 acgagcactt cttcatcgtg gaccgcctga agtctctgat caagtacaag ggctaccagg
3961 tggccccagc cgagctggag tctatcctgc tgcagcaccc taacattttc gacgccggag
4021 tggccggcct gcccgacgac gatgccggcg agctgcctgc cgccgtcgtc gtgctggaac
4081 acggcaagac catgaccgag aaggagatcg tggactatgt ggccagccag gtgacaaccg
4141 ccaagaagct gcgcggcgga gtggtgttcg tggacgaggt gcccaagggc ctgaccggca
4201 agctggacgc ccgcaagatc cgcgagatcc tgatcaaggc taagaaaggc ggcaagatcg
4261 ccgtgtaata attctagagt cggggcggcc ggccgcttcg agcagacatg ataagataca
4321 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
4381 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
4441 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
4501 agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc aggtggcact tttcggggaa
4561 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
4621 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
4681 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
4741 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
4801 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
4861 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
4921 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
4981 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
5041 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
5101 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
5161 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
5221 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
5281 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
5341 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
5401 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
5461 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
5521 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
5581 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
5641 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
5701 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
5761 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
5821 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
5881 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
5941 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
6001 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
6061 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
6121 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
6181 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
6241 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
6301 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg gctcgac
//