LOCUS Exported 7905 bp ds-DNA circular SYN 17-JAN-2017
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7905)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 28, 2017 from MiaoLingPlasmid
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..7905
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 12..467
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(560..1375)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin 1497..2085
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 2271..2413
/label=bom
/note="basis of mobility region from pBR322"
CDS complement(2515..2706)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(3507..4124)
/codon_start=1
/label=FixJ
/translation="MTTKGHIYVIDDDAAMRDSLNFLLDSAGFGVTLFDDAQAFLDALP
GLSFGCVVSDVRMPGLDGIELLKRMKAQQSPFPILIMTGHGDVPLAVEAMKLGAVDFLE
KPFEDDRLTAMIESAIRQAEPAAKSEAVAQDIAARVASLSPRERQVMEGLIAGLSNKLI
AREYDISPRTIEVYRANVMTKMQANSLSELVRLAMRAGMLND"
misc_feature complement(3507..4124)
/label=FixJ
misc_feature complement(3507..4124)
/label=FixJ
/note="YF1/FixJ bicistronic operon"
misc_feature 3510..5263
/label=YF1/FixJ
/note="bicistronic operon"
misc_feature complement(4121..5263)
/label=YF1
/note="YF1/FixJ bicistronic operon"
promoter complement(5264..5341)
/gene="lacI (mutant)"
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
misc_feature 5636..5882
/label=FixK2 promoter
RBS 5889..5900
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 5907..6617
/codon_start=1
/gene="cI"
/product="phage lambda repressor
cI-LVA"
/label=-lambda repressor
/translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
VGKVIASQWPEETFG"
terminator 6698..6769
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 6785..6812
/label=T7Te terminator
/note="phage T7 early transcription terminator"
misc_feature 6827..6875
/label=lambda pR promoter
/note="lambda pR promoter"
RBS 6888..6910
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 6929..6946
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 6956..6973
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
CDS 6977..7009
/codon_start=1
/product="leader peptide from bacteriophage T7 gene 10"
/label=T7 tag (gene 10 leader)
/note="promotes efficient translation in E. coli"
/translation="MASMTGGQQMG"
CDS 7017..7727
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
CDS 7749..7766
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
terminator 7833..7880
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
ORIGIN
1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt
481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
541 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat
601 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
661 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
721 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
781 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
841 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
901 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
961 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
1021 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
1081 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
1141 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
1201 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
1261 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
1321 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac
1381 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa
1441 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
1501 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
1561 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
1621 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
1681 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
1741 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
1801 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
1861 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
1921 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
1981 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
2041 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
2101 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
2161 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
2221 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
2281 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta
2341 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
2401 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
2461 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
2521 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
2581 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
2641 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
2701 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
2761 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
2821 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
2881 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
2941 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
3001 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
3061 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
3121 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
3181 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
3241 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
3301 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
3361 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
3421 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
3481 atgagtgagc taacttacat caattgtcaa tcgttgagca tgccggcgcg catcgcgagg
3541 cgaaccagct ccgaaaggct gttggcctgc atcttggtca tgacgttggc ccgatacacc
3601 tcgatggtgc gcgggctgat gtcgtactcg cgggcgatca gcttgttgga aaggccggcg
3661 atcagccctt ccatgacctg gcgctccctg gggctcaacg aggcgacgcg ggcggcgata
3721 tcctgcgcga cggcctcgct cttggcggcc ggctcggcct ggcggatcgc cgattcgatc
3781 atggcggtga ggcggtcgtc ctcgaaaggc ttttccagaa agtccaccgc ccctaacttc
3841 atcgcctcga ccgcgagcgg cacgtcgccg tgaccggtca tgatgaggat cggaaagggg
3901 ctttgctgcg ccttcatccg cttcaacagc tcgatgccgt caaggcccgg catgcgcacg
3961 tcggagacga cgcagccgaa ggagagaccc ggcagggcgt cgagaaaggc ttgcgcgtcg
4021 tcaaacagcg tgacgccgaa gccggcagaa tccagcagga aattcagcga atcccgcatc
4081 gccgcgtcgt cgtcgatgac gtagatatgt cccttggtcg tcatgctaga cctcctatca
4141 tctcgtcggc tgccgggagg gtgaagcgga aggtcgcccc gcccgatgcg ttgctctcgg
4201 cccacatgcg cccgccgtga gcttcgatga tcgagcggct gatggacagt cccacgccca
4261 tgccggtgtc cttggtggtg aagaaagtct gaaacaggtt cggaatgacg tcgtcctgga
4321 aaccgctgcc ggtgtcggac acttcgacct cgatcatgtc gtcggcggcg ggggtgttgg
4381 tgacgacgag ctcgcgtcgc tgcgactgag ccatcgcttc cagcgcgttg cggaacaggt
4441 tgaccaggac ctgctggatc tgcacccggt cggcgagaac gagatcggcg cccggatcga
4501 gactgaagcg gagctgcacg ttctgctcgc gcgcgccggc aagcccgagc gcgccggcct
4561 cctcgatcag cttggagaga ctctcgaccc gcttctccga ttcgccgcgg gcaacgaagt
4621 cgcgcaggcg ccggatgatc tggccggcgc gcagcgcctg ctcggcggcg cggtccaggg
4681 cgctttcgac cttcggtgtg ttcggatcac tgctgccggc aagcagccgc cgcgagccct
4741 tcatgtagtt gctgatcgcc gccagcggct ggttgagctc gtgcgcgagc gcggacgcca
4801 tttcgcccat ggcgctcagc ctggagacgt ggacgagctc ggattgcagt tcctggaggc
4861 gcgcctgggt ctgctggtgc tcggtgatat cattctgaat accgacaaaa tacgttttat
4921 cctctatttc cattggatca atatttaatt cattccagaa catcgttccg tcttttttgt
4981 agttttggat ctgaacggtg accggttctt tattttgtaa agcggttctg atgttgtcca
5041 cttctgcagg atctgtgtgt ttcccctgta agaagcgaca gttctttcct aaaatttcct
5101 cggtctcgta gccggtcatt tgaacaaagc cttgatttac gtagacaata ggattatctt
5161 caagtgcggg atctgtaatt accacaccga ctcgcacgtg atcaagtgct tttttgatga
5221 cttccagctg tcctggtatc ccaaatgatt gaaaactagc cacattcacc accctcaatt
5281 gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcaccat tcgatggtgt
5341 ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg
5401 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac
5461 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
5521 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
5581 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctacgcc
5641 cgtgatcctg atcaccggct atccggacga aaacatctcg acccgggccg ccgaggccgg
5701 cgtaaaagac gtggttttga agccgcttct cgacgaaaac ctgctcaagc gtatccgccg
5761 cgccatccag gaccggcctc gggcatgacc tacggggttc tacgtaaggc acccccctta
5821 agatatcgct cgaaattttc gaacctcccg ataccgcgta ccaatgcgtc atcacaacgg
5881 agtctagaaa agaggagaaa tactagatga gcacaaaaaa gaaaccatta acacaagagc
5941 agcttgagga cgcacgtcgc cttaaagcaa tttatgaaaa aaagaaaaat gaacttggct
6001 tatcccagga atctgtcgca gacaagatgg ggatggggca gtcaggcgtt ggtgctttat
6061 ttaatggcat caatgcatta aatgcttata acgccgcatt gcttgcaaaa attctcaaag
6121 ttagcgttga agaatttagc ccttcaatcg ccagagaaat ctacgagatg tatgaagcgg
6181 ttagtatgca gccgtcactt agaagtgagt atgagtaccc tgttttttct catgttcagg
6241 cagggatgtt ctcacctgag cttagaacct ttaccaaagg tgatgcggag agatgggtaa
6301 gcacaaccaa aaaagccagt gattctgcat tctggcttga ggttgaaggt aattccatga
6361 ccgcaccaac aggctccaag ccgagctttc ctgacggaat gttaattctc gttgaccctg
6421 agcaggctgt tgagccaggt gatttctgca tagccagact tgggggtgat gagtttacct
6481 tcaagaaact gatcagggat agcggtcagg tgtttttaca accactaaac ccacagtacc
6541 caatgatccc atgcaatgag agttgttccg ttgtggggaa agttatcgct agtcagtggc
6601 ctgaagagac gtttggcgct gcaaacgacg aaaactacgc tttagtagct taataacgct
6661 gatagtgcta gtgtagatcg ctactagagc caggcatcaa ataaaacgaa aggctcagtc
6721 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca
6781 cactggctca ccttcgggtg ggcctttctg cgtttatata ctagagtaac accgtgcgtg
6841 ttgactattt tacctctggc ggtgataatg gttgctctag aaataatttt gtttaacttt
6901 aagaaggaga tataccatgg gcagcagcca tcatcatcat catcacagca gcggcctggt
6961 gccgcgcggc agccatatgg ctagcatgac tggtggacag caaatgggtc ggatccatgg
7021 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
7081 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
7141 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
7201 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
7261 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
7321 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg
7381 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
7441 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
7501 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
7561 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
7621 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
7681 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtgaaag cttgcggccg
7741 cactcgagca ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag
7801 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
7861 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggat
//