LOCUS       pJJF152_Phsp_16_        7899 bp ds-DNA     circular     11-JAN-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pJJF152_Phsp-16_48_Cas9_unc-54UTR
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7899)
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     CDS             complement(239..307)
                     /note="LacZ alpha"
                     /locus_tag="LacZ alpha"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     378..395
                     /note="M13-fwd"
                     /locus_tag="M13-fwd"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    396..401
                     /note="EcoRI"
                     /locus_tag="EcoRI"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    402..407
                     /note="SpeI"
                     /locus_tag="SpeI"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    408..757
                     /note="Phsp-16_48 (from pMB67)"
                     /locus_tag="Phsp-16_48 (from pMB67)"
                     /ApEinfo_fwdcolor="#00ffff"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    758..763
                     /note="NheI"
                     /locus_tag="NheI"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             764..4954
                     /note="Cas9 ORF with intervening intron (friedland et al)"
                     /locus_tag="Cas9 ORF with intervening intron (from
                     Friedland et al)"
                     /ApEinfo_fwdcolor="#ff8040"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     intron          2840..2890
                     /note="intron sequence"
                     /locus_tag="intron sequence"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_signal     4919..4954
                     /note="SV40 NLS"
                     /locus_tag="SV40 NLS"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    4955..4960
                     /note="AvrII"
                     /locus_tag="AvrII"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    4961..5659
                     /note="unc-54"
                     /locus_tag="unc-54 3' UTR"
                     /ApEinfo_fwdcolor="#808040"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(5674..5694)
                     /note="M13-rev"
                     /locus_tag="M13-rev"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_binding    complement(5700..5722)
                     /note="LacO"
                     /locus_tag="LacO"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(5709..5730)
                     /note="For. seq. primer prom (oBT100)"
                     /locus_tag="For. seq. primer prom (oBT100)"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     rep_origin      complement(6062..6744)
                     /direction=LEFT
                     /note="ColE1 origin"
                     /locus_tag="ColE1 origin"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     rep_origin      6065..6679
                     /note="pMB1 Replication origin"
                     /locus_tag="pMB1 Replication origin"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    6839..7699
                     /note="AmpR"
                     /locus_tag="AmpR"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             complement(6842..7501)
                     /note="AmpR(1)"
                     /locus_tag="AmpR(1)"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cactagtatg attgtagttt
      421 gaagatttca caattagagt gaatgttgtt tggttcggtt ttgtcactgt atttatactc
      481 atttccacct ttttctagaa cattcgagct gcttcttgca aaaggagggc gactcacatt
      541 cagaacattg agaaatagtg tgcgtactga agaaacccag atactttttc aatctgcgtc
      601 tctttgcacc tatggggtgt attttgaaat gaatgcatct aggaccttct agaacattct
      661 aaacggctgc aggatacggg tatataagcc aatcgtgttc agaggaaacc aatacacttt
      721 gttcaagtgc ttactgttca ttctctaaac ttcaagagct agcatggaca agaagtactc
      781 catcggactc gacatcggaa ccaactccgt cggatgggcc gtcatcaccg acgagtacaa
      841 ggtcccatcc aagaagttca aggtcctcgg aaacaccgac cgtcactcca tcaagaagaa
      901 cctcatcgga gccctcctct tcgactccgg agagaccgcc gaggccaccc gtctcaagcg
      961 taccgcccgt cgtcgttaca cccgtcgtaa gaaccgtatc tgctacctcc aggagatctt
     1021 ctccaacgag atggccaagg tcgacgactc cttcttccac cgtctcgagg agtccttcct
     1081 cgtcgaggag gacaagaagc acgagcgtca cccaatcttc ggaaacatcg tcgacgaggt
     1141 cgcctaccac gagaagtacc caaccatcta ccacctccgt aagaagctcg tcgactccac
     1201 cgacaaggcc gacctccgtc tcatctacct cgccctcgcc cacatgatca agttccgtgg
     1261 acacttcctc atcgagggag acctcaaccc agacaactcc gacgtcgaca agctcttcat
     1321 ccagctcgtc cagacctaca accagctctt cgaggagaac ccaatcaacg cctccggagt
     1381 cgacgccaag gccatcctct ccgcccgtct ctccaagtcc cgtcgtctcg agaacctcat
     1441 cgcccagctc ccaggagaga agaagaacgg actcttcgga aacctcatcg ccctctccct
     1501 cggactcacc ccaaacttca agtccaactt cgacctcgcc gaggacgcca agctccagct
     1561 ctccaaggac acctacgacg acgacctcga caacctcctc gcccagatcg gagaccagta
     1621 cgccgacctc ttcctcgccg ccaagaacct ctccgacgcc atcctcctct ccgacatcct
     1681 ccgtgtcaac accgagatca ccaaggcccc actctccgcc tccatgatca agcgttacga
     1741 cgagcaccac caggacctca ccctcctcaa ggccctcgtc cgtcagcagc tcccagagaa
     1801 gtacaaggag atcttcttcg accagtccaa gaacggatac gccggataca tcgacggagg
     1861 agcctcccag gaggagttct acaagttcat caagccaatc ctcgagaaga tggacggaac
     1921 cgaggagctc ctcgtcaagc tcaaccgtga ggacctcctc cgtaagcagc gtaccttcga
     1981 caacggatcc atcccacacc agatccacct cggagagctc cacgccatcc tccgtcgtca
     2041 ggaggacttc tacccattcc tcaaggacaa ccgtgagaag atcgagaaga tcctcacctt
     2101 ccgtatccca tactacgtcg gaccactcgc ccgtggaaac tcccgtttcg cctggatgac
     2161 ccgtaagtcc gaggagacca tcaccccatg gaacttcgag gaggtcgtcg acaagggagc
     2221 ctccgcccag tccttcatcg agcgtatgac caacttcgac aagaacctcc caaacgagaa
     2281 ggtcctccca aagcactccc tcctctacga gtacttcacc gtctacaacg agctcaccaa
     2341 ggtcaagtac gtcaccgagg gaatgcgtaa gccagccttc ctctccggag agcagaagaa
     2401 ggccatcgtc gacctcctct tcaagaccaa ccgtaaggtc accgtcaagc agctcaagga
     2461 ggactacttc aagaagatcg agtgcttcga ctccgtcgag atctccggag tcgaggaccg
     2521 tttcaacgcc tccctcggaa cctaccacga cctcctcaag atcatcaagg acaaggactt
     2581 cctcgacaac gaggagaacg aggacatcct cgaggacatc gtcctcaccc tcaccctctt
     2641 cgaggaccgt gagatgatcg aggagcgtct caagacctac gcccacctct tcgacgacaa
     2701 ggtcatgaag cagctcaagc gtcgtcgtta caccggatgg ggacgtctct cccgtaagct
     2761 catcaacgga atccgtgaca agcagtccgg aaagaccatc ctcgacttcc tcaagtccga
     2821 cggattcgcc aaccgtaacg taagtttaaa catgatttta ctaactaact aatctgattt
     2881 aaattttcag ttcatgcagc tcatccacga cgactccctc accttcaagg aggacatcca
     2941 gaaggcccag gtctccggac agggagactc cctccacgag cacatcgcca acctcgccgg
     3001 atccccagcc atcaagaagg gaatcctcca gaccgtcaag gtcgtcgacg agctcgtcaa
     3061 ggtcatggga cgtcacaagc cagagaacat cgtcatcgag atggcccgtg agaaccagac
     3121 cacccagaag ggacagaaga actcccgtga gcgtatgaag cgtatcgagg agggaatcaa
     3181 ggagctcgga tcccagatcc tcaaggagca cccagtcgag aacacccagc tccagaacga
     3241 gaagctctac ctctactacc tccagaacgg acgtgacatg tacgtcgacc aggagctcga
     3301 catcaaccgt ctctccgact acgacgtcga ccacatcgtc ccacagtcct tcctcaagga
     3361 cgactccatc gacaacaagg tcctcacccg ttccgacaag aaccgtggaa agtccgacaa
     3421 cgtcccatcc gaggaggtcg tcaagaagat gaagaactac tggcgtcagc tcctcaacgc
     3481 caagctcatc acccagcgta agttcgacaa cctcaccaag gccgagcgtg gaggactctc
     3541 cgagctcgac aaggccggat tcatcaagcg tcagctcgtc gagacccgtc agatcaccaa
     3601 gcacgtcgcc cagatcctcg actcccgtat gaacaccaag tacgacgaga acgacaagct
     3661 catccgtgag gtcaaggtca tcaccctcaa gtccaagctc gtctccgact tccgtaagga
     3721 cttccagttc tacaaggtcc gtgagatcaa caactaccac cacgcccacg acgcctacct
     3781 caacgccgtc gtcggaaccg ccctcatcaa gaagtaccca aagctcgagt ccgagttcgt
     3841 ctacggagac tacaaggtct acgacgtccg taagatgatc gccaagtccg agcaggagat
     3901 cggaaaggcc accgccaagt acttcttcta ctccaacatc atgaacttct tcaagaccga
     3961 gatcaccctc gccaacggag agatccgtaa gcgtccactc atcgagacca acggagagac
     4021 cggagagatc gtctgggaca agggacgtga cttcgccacc gtccgtaagg tcctctccat
     4081 gccacaggtc aacatcgtca agaagaccga ggtccagacc ggaggattct ccaaggagtc
     4141 catcctccca aagcgtaact ccgacaagct catcgcccgt aagaaggact gggacccaaa
     4201 gaagtacgga ggattcgact ccccaaccgt cgcctactcc gtcctcgtcg tcgccaaggt
     4261 cgagaaggga aagtccaaga agctcaagtc cgtcaaggag ctcctcggaa tcaccatcat
     4321 ggagcgttcc tccttcgaga agaacccaat cgacttcctc gaggccaagg gatacaagga
     4381 ggtcaagaag gacctcatca tcaagctccc aaagtactcc ctcttcgagc tcgagaacgg
     4441 acgtaagcgt atgctcgcct ccgccggaga gctccagaag ggaaacgagc tcgccctccc
     4501 atccaagtac gtcaacttcc tctacctcgc ctcccactac gagaagctca agggatcccc
     4561 agaggacaac gagcagaagc agctcttcgt cgagcagcac aagcactacc tcgacgagat
     4621 catcgagcag atctccgagt tctccaagcg tgtcatcctc gccgacgcca acctcgacaa
     4681 ggtcctctcc gcctacaaca agcaccgtga caagccaatc cgtgagcagg ccgagaacat
     4741 catccacctc ttcaccctca ccaacctcgg agccccagcc gccttcaagt acttcgacac
     4801 caccatcgac cgtaagcgtt acacctccac caaggaggtc ctcgacgcca ccctcatcca
     4861 ccagtccatc accggactct acgagacccg tatcgacctc tcccagctcg gaggagactc
     4921 ccgtgccgac ccaaagaaga agcgtaaggt ctgacctagg gtccaattac tcttcaacat
     4981 ccctacatgc tctttctccc tgtgctccca ccccctattt ttgttattat caaaaaactt
     5041 ctcttaattt ctttgttttt tagcttcttt taagtcacct ctaacaatga aattgtgtag
     5101 attcaaaaat agaattaatt cgtaataaaa agtcgaaaaa aattgtgctc cctcccccca
     5161 ttaataataa ttctatccca aaatctacac aatgttctgt gtacacttct tatgtttttt
     5221 acttctgata aatttttttg aaacatcata gaaaaaaccg cacacaaaat accttatcat
     5281 atgttacgtt tcagtttatg accgcaattt ttatttcttc gcacgtctgg gcctctcatg
     5341 acgtcaaatc atgctcatcg tgaaaaagtt ttggagtatt tttggaattt ttcaatcaag
     5401 tgaaagttta tgaaattaat tttcctgctt ttgctttttg gggtttcccc tattgtttgt
     5461 caagatttcg aggacggcgt ttttcttgct aaaatcacaa gtattgatga gcacgatgca
     5521 agaaagatcg gaagaaggtt tgggtttgag gctcagtgga aggtgagtag aagttgataa
     5581 tttgaaagtg gagtagtgtc tatggggttt ttgccttaaa tgacagaata cattcccaat
     5641 ataccaaaca taactgttta agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5701 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5761 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5821 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5881 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5941 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6001 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6061 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6121 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6181 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6241 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6301 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6361 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6421 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6481 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     6541 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6601 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6661 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6721 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6781 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6841 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6901 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6961 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7021 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7081 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7141 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7201 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7261 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7321 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7381 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7441 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7501 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7561 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7621 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7681 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7741 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7801 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     7861 taacctataa aaataggcgt atcacgaggc cctttcgtc  
//
