LOCUS Exported 3515 bp ds-DNA circular SYN 30-NOV-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..3515
/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"
promoter 696..936
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
misc_RNA 963..1038
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
rep_origin complement(1535..2123)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2200..2994)
/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(3029..3386)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3043..3178
/label=SV40 ori
/note="SV40 origin of replication"
promoter complement(3388..3492)
/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 tgggtacccg
661 tctcacaggc ggatcgatcc aaggtcgggc aggaagaggg cctatttccc atgattcctt
721 catatttgca tatacgatac aaggctgtta gagagataat tggaattaat ttgactgtaa
781 acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg
841 cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt
901 tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccgggtct tcgagaagac
961 ctgttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa
1021 gtggcaccga gtcggtgctt tttttctccg ctgagcgtac tgagacgccg cggtggagct
1081 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
1141 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
1201 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
1261 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
1321 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
1381 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
1441 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
1501 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
1561 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
1621 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
1681 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
1741 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
1801 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
1861 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
1921 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
1981 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
2041 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
2101 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
2161 agtggaacga aaactcacgt taagggattt tggtcatgat cagaagaact cgtcaagaag
2221 gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg
2281 gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg
2341 atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc
2401 caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg
2461 catgctcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc
2521 cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg
2581 tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc
2641 atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc
2701 cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc
2761 tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cttgcagttc
2821 attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag
2881 ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag
2941 cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa
3001 cgatcctcat cctgtctctt gatcgatctt tgcaaaagcc taggcctcca aaaaagcctc
3061 ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa
3121 aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg
3181 gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc
3241 tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc
3301 tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat
3361 tccacagctg gttctttccg cctcaggact cttccttttt caatattatt gaagcattta
3421 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
3481 aggggttccg cgcacatttc cccgaaaagt gccac
//