LOCUS Exported 6050 bp ds-DNA circular SYN 12-JUL-2017
DEFINITION synthetic circular DNA
KEYWORDS pET28a-EGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6050)
TITLE Direct Submission
JOURNAL Exported 2015-4-17 from MLCC http://www.miaolingbio.com
REFERENCE 2 (bases 1 to 6050)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Wednesday, July 12, 2017 from miaoling plasmid
http://www.miaoligbio.com
FEATURES Location/Qualifiers
source 1..6050
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 12..467
/direction=RIGHT
/note="f1 ori"
/note="f1 ori
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"
/note="aph(3')-Ia"
/note="KanR
confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin 1497..2085
/direction=RIGHT
/note="ori"
/note="ori
high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 2271..2413
/note="bom"
/note="bom
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"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(3515..4597)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/note="lacI"
/note="lacI
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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
promoter complement(4598..4675)
/gene="lacI"
/note="lacI lacI promoter"
/note="lacI promoter"
promoter 4984..5002
/note="T7 promoter"
/note="T7 promoter
promoter for bacteriophage T7 RNA polymerase"
CDS 5083..5100
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS 5131..5847
/codon_start=1
/product="enhanced GFP"
/note="enhanced GFP"
/note="EGFP
mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
misc_feature 5848..5893
/note="MCS"
CDS 5894..5911
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
terminator 5978..6025
/note="T7 terminator"
/note="T7 terminator
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 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
3541 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
3601 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
3661 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
3721 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
3781 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
3841 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
3901 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
3961 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
4021 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
4081 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
4141 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
4201 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
4261 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
4321 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
4381 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
4441 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
4501 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
4561 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
4621 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
4681 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
4741 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc
4801 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
4861 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
4921 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga
4981 aattaatacg actcactata ggggaattgt gagcggataa caattcccct ctagaaataa
5041 ttttgtttaa ctttaagaag gagatatacc atgggcagca gccatcatca tcatcatcac
5101 agcagcggcc tggtgccgcg cggcagccat atggtgagca agggcgagga gctgttcacc
5161 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
5221 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
5281 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
5341 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
5401 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
5461 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
5521 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
5581 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
5641 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
5701 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
5761 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
5821 actctcggca tggacgagct gtacaaggga tccgaattcg agctccgtcg acaagcttgc
5881 ggccgcactc gagcaccacc accaccacca ctgagatccg gctgctaaca aagcccgaaa
5941 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
6001 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat
