LOCUS Exported 6097 bp ds-DNA circular SYN 24-SEP-2017
DEFINITION synthetic circular DNA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6097)
TITLE Direct Submission
JOURNAL Exported Sep 24, 2017 from MiaoLingPlasmid
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..6097
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 135..166
/label=minP
/note="minimal TATA-box promoter with low basal activity"
CDS 228..1877
/codon_start=1
/product="firefly luciferase"
/label=luciferase
/note="synthetic luc2 version of the luciferase gene"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
CDS 1881..2000
/codon_start=1
/product="PEST degradation sequence from mouse ornithine
decarboxylase"
/label=hPEST
/note="human codon-optimized"
/translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
polyA_signal 2052..2173
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter 2367..2724
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2575..2710
/label=SV40 ori
/note="SV40 origin of replication"
CDS 2755..3792
/codon_start=1
/product="hygromycin B phosphotransferase"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
EVGRV"
polyA_signal 3816..3864
/note="synthetic polyadenylation signal"
rep_origin complement(4191..4779)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4979..5839)
/codon_start=1
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
polyA_signal 5944..5992
/note="synthetic polyadenylation signal"
misc_feature 6006..6097
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
ORIGIN
1 ggcctaactg gccggtacct gagctcgcta gcggaggaaa aactgtttca tacagaaggc
61 gtggaggaaa aactgtttca tacagaaggc gtggaggaaa aactgtttca tacagaaggc
121 gtagatctag actctagagg gtatataatg gaagctcgaa ttccagcttg gcattccggt
181 actgttggta aaaagcttgg caatccggta ctgttggtaa agccaccatg gaagatgcca
241 aaaacattaa gaagggccca gcgccattct acccactcga agacgggacc gccggcgagc
301 agctgcacaa agccatgaag cgctacgccc tggtgcccgg caccatcgcc tttaccgacg
361 cacatatcga ggtggacatt acctacgccg agtacttcga gatgagcgtt cggctggcag
421 aagctatgaa gcgctatggg ctgaatacaa accatcggat cgtggtgtgc agcgagaata
481 gcttgcagtt cttcatgccc gtgttgggtg ccctgttcat cggtgtggct gtggccccag
541 ctaacgacat ctacaacgag cgcgagctgc tgaacagcat gggcatcagc cagcccaccg
601 tcgtattcgt gagcaagaaa gggctgcaaa agatcctcaa cgtgcaaaag aagctaccga
661 tcatacaaaa gatcatcatc atggatagca agaccgacta ccagggcttc caaagcatgt
721 acaccttcgt gacttcccat ttgccacccg gcttcaacga gtacgacttc gtgcccgaga
781 gcttcgaccg ggacaaaacc atcgccctga tcatgaacag tagtggcagt accggattgc
841 ccaagggcgt agccctaccg caccgcaccg cttgtgtccg attcagtcat gcccgcgacc
901 ccatcttcgg caaccagatc atccccgaca ccgctatcct cagcgtggtg ccatttcacc
961 acggcttcgg catgttcacc acgctgggct acttgatctg cggctttcgg gtcgtgctca
1021 tgtaccgctt cgaggaggag ctattcttgc gcagcttgca agactataag attcaatctg
1081 ccctgctggt gcccacacta tttagcttct tcgctaagag cactctcatc gacaagtacg
1141 acctaagcaa cttgcacgag atcgccagcg gcggggcgcc gctcagcaag gaggtaggtg
1201 aggccgtggc caaacgcttc cacctaccag gcatccgcca gggctacggc ctgacagaaa
1261 caaccagcgc cattctgatc acccccgaag gggacgacaa gcctggcgca gtaggcaagg
1321 tggtgccctt cttcgaggct aaggtggtgg acttggacac cggtaagaca ctgggtgtga
1381 accagcgcgg cgagctgtgc gtccgtggcc ccatgatcat gagcggctac gttaacaacc
1441 ccgaggctac aaacgctctc atcgacaagg acggctggct gcacagcggc gacatcgcct
1501 actgggacga ggacgagcac ttcttcatcg tggaccggct gaagagcctg atcaaataca
1561 agggctacca ggtagcccca gccgaactgg agagcatcct gctgcaacac cccaacatct
1621 tcgacgccgg ggtcgccggc ctgcccgacg acgatgccgg cgagctgccc gccgcagtcg
1681 tcgtgctgga acacggtaaa accatgaccg agaaggagat cgtggactat gtggccagcc
1741 aggttacaac cgccaagaag ctgcgcggtg gtgttgtgtt cgtggacgag gtgcctaaag
1801 gactgaccgg caagttggac gcccgcaaga tccgcgagat tctcattaag gccaagaagg
1861 gcggcaagat cgccgtgaat tctcacggct tccctcccga ggtggaggag caggccgccg
1921 gcaccctgcc catgagctgc gcccaggaga gcggcatgga tagacaccct gctgcttgcg
1981 ccagcgccag gatcaacgtc taaggccgcg actctagagt cggggcggcc ggccgcttcg
2041 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
2101 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
2161 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
2221 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc
2281 gtttgcgtat tgggcgctct tccgctgatc tgcgcagcac catggcctga aataacctct
2341 gaaagaggaa cttggttagc taccttctga ggcggaaaga accagctgtg gaatgtgtgt
2401 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
2461 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
2521 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
2581 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
2641 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
2701 tttggaggcc taggcttttg caaaaagctc gattcttctg acactagcgc caccatgaag
2761 aagcccgaac tcaccgctac cagcgttgaa aaatttctca tcgagaagtt cgacagtgtg
2821 agcgacctga tgcagttgtc ggagggcgaa gagagccgag ccttcagctt cgatgtcggc
2881 ggacgcggct atgtactgcg ggtgaatagc tgcgctgatg gcttctacaa agaccgctac
2941 gtgtaccgcc acttcgccag cgctgcacta cccatccccg aagtgttgga catcggcgag
3001 ttcagcgaga gcctgacata ctgcatcagt agacgcgccc aaggcgttac tctccaagac
3061 ctccccgaaa cagagctgcc tgctgtgtta cagcctgtcg ccgaagctat ggatgctatt
3121 gccgccgccg acctcagtca aaccagcggc ttcggcccat tcgggcccca aggcatcggc
3181 cagtacacaa cctggcggga tttcatttgc gccattgctg atccccatgt ctaccactgg
3241 cagaccgtga tggacgacac cgtgtccgcc agcgtagctc aagccctgga cgaactgatg
3301 ctgtgggccg aagactgtcc cgaggtgcgc cacctcgtcc atgccgactt cggcagcaac
3361 aacgtcctga ccgacaacgg ccgcatcacc gccgtaatcg actggtccga agctatgttc
3421 ggggacagtc agtacgaggt ggccaacatc ttcttctggc ggccctggct ggcttgcatg
3481 gagcagcaga ctcgctactt cgagcgccgg catcccgagc tggccggcag ccctcgtctg
3541 cgagcctaca tgctgcgcat cggcctggat cagctctacc agagcctcgt ggacggcaac
3601 ttcgacgatg ctgcctgggc tcaaggccgc tgcgatgcca tcgtccgcag cggggccggc
3661 accgtcggtc gcacacaaat cgctcgccgg agcgcagccg tatggaccga cggctgcgtc
3721 gaggtgctgg ccgacagcgg caaccgccgg cccagtacac gaccgcgcgc taaggaggta
3781 ggtcgagttt aaactctaga accggtcatg gccgcaataa aatatcttta ttttcattac
3841 atctgtgtgt tggttttttg tgtgttcgaa ctagatgctg tcgaccgatg cccttgagag
3901 ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta
3961 tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc
4021 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
4081 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
4141 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
4201 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
4261 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
4321 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
4381 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
4441 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
4501 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
4561 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
4621 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
4681 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
4741 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
4801 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
4861 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
4921 agtatatatg agtaaacttg gtctgacagc ggccgcaaat gctaaaccac tgcagtggtt
4981 accagtgctt gatcagtgag gcaccgatct cagcgatctg cctatttcgt tcgtccatag
5041 tggcctgact ccccgtcgtg tagatcacta cgattcgtga gggcttacca tcaggcccca
5101 gcgcagcaat gatgccgcga gagccgcgtt caccggcccc cgatttgtca gcaatgaacc
5161 agccagcagg gagggccgag cgaagaagtg gtcctgctac tttgtccgcc tccatccagt
5221 ctatgagctg ctgtcgtgat gctagagtaa gaagttcgcc agtgagtagt ttccgaagag
5281 ttgtggccat tgctactggc atcgtggtat cacgctcgtc gttcggtatg gcttcgttca
5341 actctggttc ccagcggtca agccgggtca catgatcacc catattatga agaaatgcag
5401 tcagctcctt agggcctccg atcgttgtca gaagtaagtt ggccgcggtg ttgtcgctca
5461 tggtaatggc agcactacac aattctctta ccgtcatgcc atccgtaaga tgcttttccg
5521 tgaccggcga gtactcaacc aagtcgtttt gtgagtagtg tatacggcga ccaagctgct
5581 cttgcccggc gtctatacgg gacaacaccg cgccacatag cagtactttg aaagtgctca
5641 tcatcgggaa tcgttcttcg gggcggaaag actcaaggat cttgccgcta ttgagatcca
5701 gttcgatata gcccactctt gcacccagtt gatcttcagc atcttttact ttcaccagcg
5761 tttcggggtg tgcaaaaaca ggcaagcaaa atgccgcaaa gaagggaatg agtgcgacac
5821 gaaaatgttg gatgctcata ctcgtccttt ttcaatatta ttgaagcatt tatcagggtt
5881 actagtacgt ctctcaagga taagtaagta atattaaggt acgggaggta ttggacaggc
5941 cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata
6001 gtactaacat acgctctcca tcaaaacaaa acgaaacaaa acaaactagc aaaataggct
6061 gtccccagtg caagtgcagg tgccagaaca tttctct
//