LOCUS       sgPlxnb2-3_in_pT4-U6-sg 5399 bp ds-DNA     circular     02-JUN-2022
DEFINITION  .
FEATURES             Location/Qualifiers
     CDS             complement(138..206)
                     /label="LacZ alpha"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     primer_bind     277..294
                     /label="M13-fwd"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer_bind     303..323
                     /label="T7"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    complement(348..591)
                     /label="pT4 LIR"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer          582..618
                     /label="InF-U6-F"
                     /note="sequence: ctgtgcctttaagcttggtaccgagggcctatttccc"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     primer          604..624
                     /label="hU6-f_Microsynth"
                     /note="sequence: GAGGGCCTATTTCCCATGATT"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    604..852
                     /label="U6 promoter"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer          773..797
                     /label="qPCR-screen-U6-F2"
                     /note="sequence: tggactatcatatgcttaccgtaac"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer          835..852
                     /label="qPCR-screen-U6-F"
                     /note="sequence: tggaaaggacgaaacacc"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer          849..872
                     /label="sgPlxnb2-3 FWD"
                     /note="sequence: caccgggaaacgcggggtacgcta"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    853..872
                     /label="Guide"
                     /ApEinfo_revcolor="#7ea7d8"
                     /ApEinfo_fwdcolor="#7ea7d8"
     primer          complement(853..876)
                     /label="sgPlxnb2-3 REV"
                     /note="sequence: aaactagcgtaccccgcgtttccc"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_feature    873..954
                     /label="chimeric RNA backbone 1 (BsmBI)"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          complement(934..970)
                     /label="InF-sgRNA-Hind-R"
                     /note="sequence: aatcaatgccaagcttaaaaaagcaccgactcggtgc"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     misc_feature    961..1340
                     /label="CMV enhancer"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     misc_feature    1341..1544
                     /label="CMV promoter"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     misc_feature    1636..1768
                     /label="chimeric intron"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     misc_feature    1773..1782
                     /label="kozak"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    1779..2498
                     /label="EGFP CDS"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     misc_feature    2505..2553
                     /label="synthetic poly(A) signal"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_feature    2591..2856
                     /label="pT4 RIR"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     primer_bind     complement(2887..2906)
                     /label="T3"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     promoter        complement(2889..2907)
                     /label="T3 promoter"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer_bind     complement(2924..2944)
                     /label="M13-rev"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_binding    complement(2950..2972)
                     /label="LacO"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     promoter        complement(2976..3006)
                     /label="lac promoter"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_feature    complement(3330..3918)
                     /label="Ori"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    complement(4089..4949)
                     /label="AmpR"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
ORIGIN
        1 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
       61 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
      121 attaacgctt acaatttcca ttcgccattc aggctgcgca actgttggga agggcgatcg
      181 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
      241 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc
      301 gcgtaatacg actcactata gggcgaattg gagctagcgt ttaaacctac agttgaagtc
      361 ggaagtttac atacacttaa gttggagtca ttaaaactcg tttttcaact actccacaaa
      421 tttcttgtta acaaacaata gttttggcaa gtcagttagg acatctactt tgtgcatgac
      481 acaagtcatt tttccaacaa ttgtttacag acagattatt tcacttataa ttcactgtat
      541 cacaattcca gtgggtcaga agtgtacata cacgcgcttg actgtgcctt taagcttggt
      601 accgagggcc tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga
      661 gagataatta gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt
      721 agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat
      781 catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttgtggaaa
      841 ggacgaaaca ccgggaaacg cggggtacgc tagttttaga gctagaaata gcaagttaaa
      901 ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt ttttaagctt
      961 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     1021 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     1081 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     1141 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     1201 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     1261 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
     1321 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
     1381 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     1441 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
     1501 tgggcggtag gcgtgtacgg tgggaggtct atataagcag aggtcgttta gtgaaccgtc
     1561 agatcactag tagctttatt gcggtagttt atcacagtta aattgctaac gcagtcagtg
     1621 ctcgactgat cacaggtaag tatcaaggtt acaagacagg tttaaggagg ccaatagaaa
     1681 ctgggcttgt cgagacagag aagattcttg cgtttctgat aggcacctat tggtcttact
     1741 gacatccact ttgcctttct ctccacaggg cggccgccat ggtgagcaag ggcgaggagc
     1801 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     1861 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     1921 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     1981 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     2041 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     2101 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     2161 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     2221 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     2281 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     2341 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     2401 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     2461 ccgggatcac tctcggcatg gacgagctgt acaagtaaga attcaataaa atatctttat
     2521 tttcattaca tctgtgtgtt ggttttttgt gtgtaattcg agcatcttac cgccatttat
     2581 tcccagatct ttaaacaatt taaaggcaat gctaccaaat actaagcgcg tgtatgtaca
     2641 cttctgaccc actgggaatg tgatgaaaga aataaaagct gaaatgaatc attctctcta
     2701 ctattattct gatatttcac attcttaaaa taaagtggtg atcctaactg accttaagac
     2761 agggaatctt tactcggatt aaatgtcagg aattgtgaaa aagtgagttt aaatgtattt
     2821 ggctaaggtg tatgtaaact tccgacttca actgtagttt aaacgggccc ggtacccagc
     2881 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
     2941 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     3001 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     3061 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     3121 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3181 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3241 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3301 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3361 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3421 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3481 tacctgtccg cctttctccc ttcgggaacc gtggcgcttt ctcatagctc acgctgtagg
     3541 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3601 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3661 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3721 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3781 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3841 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3901 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3961 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4021 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4081 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4141 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4201 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4261 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4321 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4381 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4441 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4501 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaactt ggccgcagtg
     4561 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4621 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4681 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4741 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4801 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4861 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4921 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     4981 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5041 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca
     5101 ttaagcgcgg cgggagtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     5161 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     5221 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     5281 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     5341 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgg
//