LOCUS Exported 3451 bp ds-DNA circular SYN 07-JAN-2018
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..3451
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..215
/label=H1 promoter
/note="human H1 RNA promoter"
misc_RNA 242..317
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
rep_origin complement(803..1391)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1468..2262)
/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(2297..2654)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2311..2446
/label=SV40 ori
/note="SV40 origin of replication"
promoter complement(2656..2760)
/gene="bla"
/label=AmpR promoter
rep_origin complement(2786..3241)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
ORIGIN
1 gaacgctgac gtcatcaacc cgctccaagg aatcgcgggc ccagtgtcac taggcgggaa
61 cacccagcgc gcgtgcgccc tggcaggaag atggctgtga gggacagggg agtggcgccc
121 tgcaatattt gcatgtcgct atgtgttctg ggaaatcacc ataaacgtga aatgtctttg
181 gatttgggaa tcttataagt tctgtatgag accactcttt cccaggtctt cgagaagacc
241 tgttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag
301 tggcaccgag tcggtgcttt ttttggtctg agacgccgcg gtggagctcc agcttttgtt
361 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
421 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
481 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
541 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
601 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
661 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
721 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
781 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
841 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
901 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
961 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
1021 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
1081 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
1141 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
1201 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
1261 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
1321 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
1381 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
1441 actcacgtta agggattttg gtcatgatca gaagaactcg tcaagaaggc gatagaaggc
1501 gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc
1561 gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc
1621 cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt
1681 cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt
1741 gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg
1801 atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg
1861 gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat
1921 ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc
1981 caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac
2041 gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc
2101 ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc
2161 ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca
2221 agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc
2281 tgtctcttga tcgatctttg caaaagccta ggcctccaaa aaagcctcct cactacttct
2341 ggaatagctc agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc
2401 catggggcgg agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg
2461 gcgggactat ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg
2521 agcctgggga ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc
2581 tgcctgctgg ggagcctggg gactttccac accctaactg acacacattc cacagctggt
2641 tctttccgcc tcaggactct tcctttttca atattattga agcatttatc agggttattg
2701 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
2761 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
2821 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
2881 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
2941 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
3001 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
3061 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
3121 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
3181 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
3241 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
3301 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
3361 gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta atacgactca
3421 ctatagggcg aattgggtac ccgtctcacc t
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