LOCUS Exported 5484 bp ds-DNA circular SYN 20-NOV-2016
DEFINITION synthetic circular DNA
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..5484
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 235..614
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 863..881
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 898..924
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/note="HA"
/translation="YPYDVPDYA"
misc_feature 927..1032
/note="MCS"
promoter complement(1036..1054)
/note="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
polyA_signal 1080..1304
/note="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 1350..1778
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 1792..2122
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 1973..2108
/note="SV40 ori"
/note="SV40 origin of replication"
CDS 2189..2983
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/note="NeoR"
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
polyA_signal 3157..3278
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
primer_bind complement(3327..3343)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3351..3367
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/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(3375..3405)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 3420..3441
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(3729..4317)
/direction=LEFT
/note="pUC ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4488..5348)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5349..5453)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccgc caccatgtac
901 ccatacgacg tcccagacta cgctggaagc ttggtaccga gctcggatcc actagtaacg
961 gccgccagtg tgctggaatt ctgcagatat ccatcacact ggcggccgct cgagcatgca
1021 tctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac
1081 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
1141 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
1201 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
1261 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
1321 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
1381 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
1441 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
1501 tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
1561 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
1621 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
1681 ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt
1741 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
1801 ttagggtgtg gaaagtcccc aggctcccca ggcaggcaga agtatgcaaa gcatgcatct
1861 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
1921 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
1981 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
2041 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
2101 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
2161 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
2221 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
2281 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
2341 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
2401 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
2461 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
2521 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
2581 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
2641 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
2701 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
2761 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
2821 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
2881 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
2941 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
3001 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
3061 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
3121 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
3181 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
3241 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
3301 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
3361 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
3421 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
3481 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
3541 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
3601 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
3661 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
3721 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
3781 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
3841 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
3901 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc
3961 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
4021 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
4081 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
4141 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
4201 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
4261 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
4321 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
4381 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
4441 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
4501 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
4561 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
4621 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
4681 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
4741 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
4801 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
4861 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
4921 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
4981 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
5041 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
5101 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
5161 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
5221 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
5281 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
5341 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
5401 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
5461 ccccgaaaag tgccacctga cgtc
//