LOCUS Exported 6159 bp ds-DNA circular SYN 27-SEP-2017
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6159)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 27, 2017 from MiaoLingPlasmid
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..6159
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 889..976
/label=MCS
CDS 977..1696
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
promoter complement(1712..1730)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
polyA_signal 1756..1980
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2026..2454
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2468..2797
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2648..2783
/label=SV40 ori
/note="SV40 origin of replication"
CDS 2864..3658
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=Neo
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
polyA_signal 3832..3953
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin complement(4404..4992)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5163..6023)
/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(6024..6128)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 ggagacccaa gcttggtacc
901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttctgcagat atccatcaca
961 ctggcggccg ctcgagatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
1021 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
1081 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
1141 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
1201 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
1261 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
1321 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
1381 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
1441 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
1501 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
1561 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
1621 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
1681 cgagctgtac aagtaatcta gagggcccta ttctatagtg tcacctaaat gctagagctc
1741 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
1801 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
1861 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
1921 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
1981 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
2041 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
2101 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
2161 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
2221 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
2281 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
2341 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
2401 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
2461 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
2521 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
2581 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
2641 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
2701 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
2761 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
2821 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
2881 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
2941 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
3001 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
3061 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
3121 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
3181 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
3241 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
3301 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
3361 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
3421 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
3481 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
3541 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
3601 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
3661 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
3721 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
3781 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
3841 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
3901 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
3961 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
4021 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
4081 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
4141 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
4201 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
4261 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
4321 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
4381 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
4441 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
4501 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
4561 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
4621 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
4681 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
4741 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
4801 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
4861 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
4921 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
4981 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
5041 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
5101 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
5161 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
5221 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
5281 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
5341 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
5401 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
5461 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
5521 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
5581 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
5641 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
5701 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
5761 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
5821 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
5881 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
5941 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
6001 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
6061 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
6121 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//