LOCUS Exported 5818 bp ds-DNA circular SYN 28-SEP-2017
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5818)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 28, 2017 from MiaoLingPlasmid
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..5818
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 473..1651
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
promoter 1668..1686
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 1713..1792
/label=MCS
CDS 1805..1846
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
CDS 1856..1873
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
polyA_signal 1902..2126
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2172..2600
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2614..2943
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2794..2929
/label=SV40 ori
/note="SV40 origin of replication"
promoter 2991..3038
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 3057..3455
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/label=Blast
/note="confers resistance to blasticidin"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
polyA_signal 3613..3734
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
protein_bind 3807..3823
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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(3831..3861)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3876..3897
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4185..4773)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4944..5804)
/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"
promoter complement(5805..91)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
61 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
121 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
181 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
241 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
301 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
361 ttgattattg actaggcttt tgcaaaaagc tttgcaaaga tggataaagt tttaaacaga
421 gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa ttggctccgg
481 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt
541 cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt
601 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc
661 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg
721 ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac
781 ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt gggagagttc
841 gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg
901 ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa
961 gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct ggcaagatag
1021 tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg ccgcgggcgg
1081 cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc
1141 accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc ctggcctcgc
1201 gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac cagttgcgtg
1261 agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg
1321 ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc cgtcctcagc
1381 cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg attagttctc
1441 gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga tggagtttcc
1501 ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt aattctcctt
1561 ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga cagtggttca
1621 aagttttttt cttccatttc aggtgtcgtg aggaattagc ttggtactaa tacgactcac
1681 tatagggaga cccaagctgg ctaggtaagc ttggtaccga gctcggatcc actagtccag
1741 tgtggtggaa ttctgcagat atccagcaca gtggcggccg ctcgagtcta gagggccctt
1801 cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta ccggtcatca
1861 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
1921 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
1981 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
2041 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
2101 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
2161 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
2221 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
2281 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
2341 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
2401 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
2461 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
2521 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
2581 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
2641 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
2701 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
2761 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
2821 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
2881 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
2941 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt
3001 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg
3061 ccaagccttt gtctcaagaa gaatccaccc tcattgaaag agcaacggct acaatcaaca
3121 gcatccccat ctctgaagac tacagcgtcg ccagcgcagc tctctctagc gacggccgca
3181 tcttcactgg tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc
3241 tgggcactgc tgctgctgcg gcagctggca acctgacttg tatcgtcgcg atcggaaatg
3301 agaacagggg catcttgagc ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc
3361 ctgggatcaa agccatagtg aaggacagtg atggacagcc gacggcagtt gggattcgtg
3421 aattgctgcc ctctggttat gtgtgggagg gctaagcact tcgtggccga ggagcaggac
3481 tgacacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
3541 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
3601 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
3661 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
3721 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
3781 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
3841 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
3901 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
3961 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
4021 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
4081 gtaatacggt tatccacaga atcaggggat aacgcaggaa agagcatgtg agcaaaaggc
4141 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
4201 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
4261 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
4321 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
4381 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
4441 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
4501 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
4561 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
4621 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
4681 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
4741 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
4801 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
4861 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
4921 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
4981 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
5041 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
5101 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
5161 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
5221 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
5281 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
5341 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
5401 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
5461 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
5521 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
5581 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
5641 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
5701 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
5761 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttc
//