LOCUS       pPS                     6593 bp    DNA     circular UNA 30-DEC-2016
DEFINITION  Gibson Assembly of 1_gBlock_site-site-scaf-BsmBI-hU6 into
            1_PmeI-HpaI_WeissmanPlasmid.
ACCESSION   urn.local...6vsu9by-y
VERSION     urn.local...6vsu9by-y
KEYWORDS    .
SOURCE      
  ORGANISM  
            .
FEATURES             Location/Qualifiers
     origin          0^1
                     /label="origin"
     promoter        241..269
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /label="AmpR promoter"
     promoter        595..1158
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /label="pU6 5'CMV promoter"
     LTR             1176..1356
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
                     /label="LTR 5'"
     misc_feature    1408..1545
                     /created_by="User"
                     /label="Psi"
     misc_feature    1512..1882
                     /created_by="User"
                     /label="Gag"
     misc_feature    2028..2269
                     /created_by="User"
                     /label="RRE"
     misc_feature    2787..2904
                     /created_by="User"
                     /label="cPPT"
     primer_bind     2913..2932
                     /Sequence="TTACAGGGACAGCAGAGATC"
                     /Hairpin_Tm="None"
                     /Tm="56.3"
                     /Self_Dimer_Tm="None"
                     /%GC="50.0"
                     /created_by="primer3"
                     /label="pHR-F"
     polyA_signal    complement(2963..3190)
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
                     /modified_by="User"
                     /label="ANTIPARALLEL - BGH pA"
     misc_feature    3184..3738
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     ligation        3184..3208
                     /label="Ligation"
     source          3209..3224
                     /created_by="User"
                     /modified_by="User"
                     /label="WeissmanPlasmid"
     misc_binding    3224..3243
                     /created_by="User"
                     /modified_by="User"
                     /label="GBC_CloningSite"
     motif           3224..3229
                     /created_by="User"
                     /label="EcoRI"
     motif           3238..3243
                     /created_by="User"
                     /label="EcoRI"
     misc_feature    3239..3738
                     /label="1_gBlock_site-site-scaf-BsmBI-hU6"
                     /note="Geneious type: Inserted sequence"
     misc_feature    <3244..>3295
                     /label="1_gBlock_site-site-scaf-BsmBI-hU6"
                     /note="Geneious type: Inserted sequence"
     source          3244..3248
                     /created_by="User"
                     /label="WeissmanPlasmid"
     misc_binding    3272..3295
                     /created_by="User"
                     /modified_by="User"
                     /label="EF1a-Puro-BFP_CloningSite"
     motif           3272..3279
                     /created_by="User"
                     /modified_by="User"
                     /label="NotI"
     motif           3288..3295
                     /created_by="User"
                     /modified_by="User"
                     /label="NotI"
     misc_RNA        complement(3319..3410)
                     /ApEinfo_fwdcolor="#ff3d4c"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /modified_by="User"
                     /label="Optimized scaffold sgRNA"
     misc_binding    3411..3463
                     /note="Filler"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /modified_by="User"
                     /label="Filler"
     motif           complement(3412..3417)
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /modified_by="User"
                     /label="BsmBI"
     motif           3453..3458
                     /ApEinfo_label="BsmBI recognition site"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /modified_by="User"
                     /label="BsmBI"
     promoter        complement(3460..3708)
                     /note="hU6"
                     /ApEinfo_fwdcolor="#c48d3c"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="hU6"
     misc_feature    3526..3531
                     /Recognition_pattern="CA^TATG"
                     /annotation_group="CATATG"
                     /label="NdeI, FauNDI"
                     /note="Geneious type: restriction site"
     ligation        3709..3738
                     /label="Ligation"
     LTR             3804..4404
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
                     /modified_by="User"
                     /label="WPRE"
     misc_feature    complement(3867..3887)
                     /Sequence="CATAGCGTAAAAGGAGCAACA"
                     /Hairpin_Tm="None"
                     /Tm="56.6"
                     /Self_Dimer_Tm="None"
                     /%GC="42.9"
                     /created_by="primer3"
                     /label="WPRE-R"
                     /note="Geneious type: primer_bind_reverse"
     LTR             4594..4608
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
                     /label="cPPT"
     LTR             4931..5111
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /label="3' LTR"
     rep_origin      5155..5837
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
                     /label="ColE1 origin"
     gene            complement(5935..1)
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
                     /label="AmpR"
ORIGIN      
        1 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
       61 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
      121 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
      181 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
      241 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
      301 cgcgcacatt tccccgaaaa gtgccacctg acgcttaagc ggtcgacgga tcgggagatc
      361 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
      421 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
      481 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
      541 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta
      601 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      661 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      721 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      781 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      841 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      901 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      961 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     1021 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     1081 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     1141 ggaggtctat ataagcagcg cgttttgcct gtactgggtc tctctggtta gaccagatct
     1201 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     1261 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     1321 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
     1381 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
     1441 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
     1501 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
     1561 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
     1621 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
     1681 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
     1741 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
     1801 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
     1861 gcaagcggcc ggccgcgctg atcttcagac ctggaggagg agatatgagg gacaattgga
     1921 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
     1981 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
     2041 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
     2101 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
     2161 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
     2221 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
     2281 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
     2341 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
     2401 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
     2461 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
     2521 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
     2581 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
     2641 acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga
     2701 gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattctgc
     2761 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     2821 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     2881 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg
     2941 ttagtaccgg gcccgctcta gaccatagag cccaccgcat ccccagcatg cctgctattg
     3001 tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac
     3061 ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca
     3121 ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa
     3181 ggcacagtcg aggctgatca gcgggtttaa acgggccctc taggaattca tgcatgcgaa
     3241 ttcaattcgc cagggttttc ccagtcacga cgcggccgca tgcatgcgcg gccgccacca
     3301 aacgtggctt gccagcccaa aaaagcaccg actcggtgcc actttttcaa gttgataacg
     3361 gactagcctt atttaaactt gctatgctgt ttccagcata gctcttaaac agagacgttg
     3421 aagatgaaac gtgtagtgtg ctcatttaca accgtctccg gtgtttcgtc ctttccacaa
     3481 gatatataaa gccaagaaat cgaaatactt tcaagttacg gtaagcatat gatagtccat
     3541 tttaaaacat aattttaaaa ctgcaaacta cccaagaaat tattactttc tacgtcacgt
     3601 attttgtact aatatctttg tgtttacagt caaattaatt ctaattatct ctctaacagc
     3661 cttgtatcgt atatgcaaat atgaaggaat catgggaaat aggccctcaa cgtcgaggga
     3721 cctaataact tcgtatagca tacattatac gaagttatac atgtttaagg gttccggttc
     3781 cactaggtac aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt
     3841 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     3901 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     3961 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     4021 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     4081 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     4141 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     4201 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     4261 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     4321 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     4381 tccctttggg ccgcctcccc gcatcgatac cgtcgacctc gatcgagacc tagaaaaaca
     4441 tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc
     4501 acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac
     4561 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     4621 aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta
     4681 cttccctgat tggcagaact acacaccagg gccagggatc agatatccac tgacctttgg
     4741 atggtgctac aagctagtac cagttgagca agagaaggta gaagaagcca atgaaggaga
     4801 gaacacccgc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt
     4861 attagagtgg aggtttgaca gccgcctagc atttcatcac atggcccgag agctgcatcc
     4921 ggactgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     4981 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     5041 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     5101 aatctctagc agcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5161 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5221 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5281 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5341 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5401 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5461 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5521 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5581 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5641 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5701 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5761 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5821 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5881 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5941 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6001 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6061 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6121 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6181 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6241 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6301 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6361 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6421 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6481 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6541 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tca
//
