LOCUS Exported 3504 bp ds-DNA circular SYN 20-DEC-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..3504
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin complement(3..458)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
misc_feature 691..933
/label=7SK promoter
misc_RNA 952..1027
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
rep_origin complement(1524..2112)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2189..2983)
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
promoter complement(3018..3375)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3032..3167
/label=SV40 ori
/note="SV40 origin of replication"
promoter complement(3377..3481)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtacctt
661 taattctagt actatgcatc gtctcattgt ctgcagtatt tagcatgccc cacccatctg
721 caaggcattc tggatagtgt caaaacagcc ggaaatcaag tccgtttatc tcaaacttta
781 gcattttggg aataaatgat atttgctatg ctggttaaat tagattttag ttaaatttcc
841 tgctgaagct ctagtacgat aagcaacttg acctaagtgt aaagttgaga cttccttcag
901 gtttatatag cttgtgcgcc gcttgggtac ctcgggtctt cgagaagacc tgttttagag
961 ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag
1021 tcggtgcttt ttttctcgga ctgatgacct gagacgccgc ggtggagctc cagcttttgt
1081 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
1141 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
1201 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
1261 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
1321 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
1381 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
1441 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
1501 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
1561 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
1621 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
1681 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
1741 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
1801 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
1861 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
1921 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
1981 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
2041 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
2101 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
2161 aactcacgtt aagggatttt ggtcatgatc agaagaactc gtcaagaagg cgatagaagg
2221 cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt
2281 cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg
2341 ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat
2401 tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgctcgcct
2461 tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct
2521 gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt
2581 ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga
2641 tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc
2701 ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa
2761 cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac
2821 cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg
2881 cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc
2941 aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc
3001 ctgtctcttg atcgatcttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc
3061 tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag
3121 ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg
3181 ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
3241 gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt
3301 ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt ccacagctgg
3361 ttctttccgc ctcaggactc ttcctttttc aatattattg aagcatttat cagggttatt
3421 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
3481 gcacatttcc ccgaaaagtg ccac
//