LOCUS       Copy_of_Copy_of_VB_pCas 6001 bp ds-DNA     circular     17-MAY-2022
DEFINITION  .
ACCESSION   urn.local...30-947jdbh                                             
FEATURES             Location/Qualifiers
     primer_bind     793..812
                     /label="T7"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     primer          922..938
                     /label="cas4 replacement FW (GA tail)"
                     /note="sequence: atccaatgcaATGGCTGAGACAGACGG"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     CDS             922..1533
                     /label="Translation 922-1533"
     CDS             922..1533
                     /label="Translation 922-1533"
     misc_feature    922..1533
                     /label="Replaced by I-U cas4"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     CDS             922..2592
                     /label="Translation 922-2592"
     primer          complement(1516..1533)
                     /label="cas4 replacement RV (GA tail)"
                     /note="sequence: cggactcgtGAGAAAGCGCACTTCATC"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     misc_feature    1534..2592
                     /label="Cas 4/1 fusion"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     primer          1596..1615
                     /label="VB cas2 del US FW"
                     /note="sequence: aaccgtgtacgtgtcggagc"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     primer          complement(1662..1683)
                     /label="K81->A mut DS RV (cas4 V-B)"
                     /note="sequence: ggtcgattccatcccatctttc"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     misc_feature    2592..2891
                     /label="Cas2"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     primer          complement(2593..2611)
                     /label="VB cas2 del US RV (BsmI tail)"
                     /note="sequence: ataGAATGCGacatatcggcgcatcgcca"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     primer          2892..2916
                     /label="VB cas2 del DS FW (BsmI tail)"
                     /note="sequence: tatGAATGCGaggattgatgtaaacttcaaatacg"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     terminator      2932..3082
                     /label="Tracr 151"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     primer          3083..3099
                     /label="VB_Leader_fw BG13572"
                     /note="sequence: ggtgggacaagggattc"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     misc_feature    3083..3215
                     /label="leader"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     misc_feature    3216..3251
                     /label="Geneious type: CRISPR Repeat Unit"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer          3234..3252
                     /label="V-B-Repeat-degenerate-Fw (BN1046,1047,1048)"
                     /note="sequence: cgatctgagaagtggcacv"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     primer          complement(3234..3252)
                     /label="Repeat Rv VB"
                     /note="sequence: agtgccacttctcagatcg"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     misc_feature    3252..3286
                     /label="Geneious type: CRISPR Spacer"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     primer          complement(3264..3286)
                     /label="Spacer acq assay RV (BN1049)"
                     /note="sequence: ggatgcatcattcgtctttttac"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     misc_feature    3287..3305
                     /label="misc_feature"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     primer          complement(3373..3392)
                     /label="VB cas2 del DS RV"
                     /note="sequence: ttcctttcgggctttgttag"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     primer_bind     complement(3377..3400)
                     /label="primer_bind"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     primer          complement(3377..3400)
                     /label="Universal reverse BN519"
                     /note="sequence: aactcagcttcctttcgggctttg"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    3412..3430
                     /label="misc_feature"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     misc_feature    3433..3473
                     /label="misc_feature"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     misc_feature    4664..4673
                     /label="misc_feature"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     rep_origin      complement(5216..5256)
                     /label="CloDF13 ori (fragment)"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    complement(5575..5595)
                     /label="misc_feature"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     CDS             complement(join(5591..6001,1..546))
                     /label="LacI"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
ORIGIN
        1 catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata
       61 tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag
      121 cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc
      181 atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg
      241 aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat
      301 gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac
      361 gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt
      421 aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat
      481 cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc
      541 cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac
      601 cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac
      661 tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg
      721 aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct
      781 gcattaggaa attaatacga ctcactatag ggagagaatt gtgagcggat aacaattccc
      841 ctgtagatta attaagcggc cgccctgcag gactcgagtt ctagaaataa ttttgtttaa
      901 ctttaagaag gagatataca tATGGCTGAG ACAGACGGGA GTATTCCTCT CATCCCGGTT
      961 CGCATGCTCA ACGAACACGT CTACTGCCCG CGACTGGCCT ATCTCATGTG GGTGCAGGGT
     1021 GAGTTCTCCC ACAATGAGTT CACGGTTGAT GGCGTTATCC GCCACCGCAG GGTCGATGCT
     1081 GGCGGCGGAG TGCTGCCCTC CGAGACCCAG GAGGATTCCA GGATACATGC CCGCTCGGTG
     1141 AGTCTCAGCT CGGAACGGCT GGGAATTACC GCCAAGATCG ATCTGGTGGA AGGGGAGGGA
     1201 GCATACGTTT CTCCTGTCGA TTACAAGCGG GGCAAACGCC CCCATGTGGC CGGCGGAGCA
     1261 TACGAGCCGG AGCGGGTTCA GCTCTGTGCC CAGGGGCTTC TTCTGCGGGA GCACGGATTT
     1321 GCCAGCGATG GCGGCGCTCT CTACTTCGTT GCCTCCCGCG AACGGGTGCC GGTTGCATTT
     1381 GATGATGAAC TGATCGGAAG AACCCTGGCC GCCATTGATG AGATGGGACG CACGGCGTTG
     1441 TCGGGCACGA TGCCCCCGCC GCTGGAGGAC AGTCCCAAAT GCCCTCGCTG CTCGCTGGTG
     1501 GGGATCTGTC TGCCGGATGA AGTGCGCTTT CTCacgagtc cggacggaga gaccgttccg
     1561 atacgagata ttgtccccag tcgagatgat ttaggaaccg tgtacgtgtc ggagcctggg
     1621 acccgggtcg gaaagcgtgg ttacgagttg accatcacgt cgaaagatgg gatggaatcg
     1681 accgttccgt taaaagatgt tcggcacatc tccctcttcg ggaacgttca gatttccacc
     1741 caattggtgc acgattgtat ggagtccggc gtcaccatca gctacttgac tgccgccggc
     1801 cgtctcgtcg ggatgaacca caatttgatt tcgaaaaacg tcttggtgcg gcgtgaacag
     1861 ttcaagcggt ttggggacga gcgtgatcga ctgacacttg cgaggtatat cgttcgcgcg
     1921 aaaattctca accagcgcac cttgcttcgt cgaaatgcgc acgggctcga caaggtcatt
     1981 ttgaaagatt tgctcgatca cgcccgacgt tccgaagaag ccacttccat tgactcgttg
     2041 ctcggcattg aagggattgc ggcgaaacag tacatgcagg cgtttgtgct gatgctgaaa
     2101 gtaaaaggcg ttcctgcggg cgaaacgctg atgaacggca gaaatcgccg cccaccgaag
     2161 gacccggtta atgcgttgct gtcacttgga tattcgttgt tgcaacggga tatgtatgcc
     2221 gccttggctg cagtgggcct ggaccccttg atgggctttt accatcgcat tgaaacgggg
     2281 cgtccgtcac tggtactcga catgatggaa ccgtttcgag cgatcatcgc ggattcggtg
     2341 gtcgttcgaa cattgaacac cggcgaaatt gcctgggatg acttttatat cgggcaggag
     2401 tcctgttcgt tgaagctgcc tggacggaaa cgattttttc aggcgtttga gcgacgcatg
     2461 cacgagaccg tgacgcatcc ggtgttcggt tacaaattga gttaccgtcg gatgctggaa
     2521 ctagaagcgc gcttcttgtc gcgatacttg atgggcgaat tgccggaata tcgcccaatt
     2581 gtcacgaggt gatggcgatg cgccgatatg tgctggtgag ctacgatatc agtgatcaaa
     2641 aacgctggcg aagggtatac aaaatcgttc gaggctatgg catgcacgtt cagtattcgg
     2701 tgttcctatg ccaactcacc gatcaggatg aggccgagtt gaagcaattg ctattggatg
     2761 tgattcatca gtcgatggat caagtcattt ttgcaaggat tggaaccgtg caggcgaaag
     2821 ctgccgagcg gaatatgtcg tgcattgggc gggacttcgt accattggat ttgaagggat
     2881 tggtattttg aaggattgat gtaaacttca aatacgcgta aaatataatc tgtctagagg
     2941 acagaatttt tcaacgggtg tgccaatggc cactttccag gtggcaaagc ccgttgagct
     3001 tctcaaaaag aacgctcgct cagtgttctg acctttcgag cgcctgttca gggcgaaaac
     3061 cctgggaggc gctcgaatca taggtgggac aagggattcg cggcgaaaac gacgggagag
     3121 aacgagaatg aaacgagatg aaatcgatgg gaaacatcaa gtgctcgttt atgggcttgg
     3181 atctccgcgt ggttcaaggg ctgcgagcgc ccgctgtcgg atcactgagc gagcgatctg
     3241 agaagtggca ctgtttggta aaggtaaaaa gacgaatgat gcatcctaac attggaagtg
     3301 gataacggat ccgcgatcgc ggcgcgccac ctggtggccg gccggtacca cgcgtgcgcg
     3361 ctgatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
     3421 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaac
     3481 ctcaggcatt tgagaagcac acggtcacac tgcttccggt agtcaataaa ccggtaaacc
     3541 agcaatagac ataagcggct atttaacgac cctgccctga accgacgacc gggtcatcgt
     3601 ggccggatct tgcggcccct cggcttgaac gaattgttag acattatttg ccgactacct
     3661 tggtgatctc gcctttcacg taatggacaa attcttccaa ctgatctgcg cgcgaggcca
     3721 agcgatcttc ttcttgtcca agataagcct gtctagcttc aagtatgacg ggctgatact
     3781 gggccggcag gcgctccatt gcccagtcgg cagcgacatc cttcggcgcg attttgccgg
     3841 ttactgcgct gtaccaaatg cgggacaacg taagcactac atttcgctca tcgccagccc
     3901 agtcgggcgg cgagttccat agcgttaagg tttcatttag cgcctcaaat agatcctgtt
     3961 caggaaccgg atcaaagagt tcctccgccg ctggacctac caaggcaacg ctatgttctc
     4021 ttgcttttgt cagcaagata gccagatcaa tgtcgatcgt ggctggctcg aagatacctg
     4081 caagaatgtc attgcgctgc cattctccaa attgcagttc gcgcttagct ggataacgcc
     4141 acggaatgat gtcgtcgtgc acaacaatgg tgacttctac agcgcggaga atctcgctct
     4201 ctccagggga agccgaagtt tccaaaaggt cgttgatcaa agctcgccgc gttgtttcat
     4261 caagccttac ggtcaccgta accagcaaat caatatcact gtgcggcttc aggccgccat
     4321 ccactgcgga gccgtacaaa tgtacggcca gcaacgtcgg ttcgagatgg cgctcgatga
     4381 cgccaactac ctctgatagt tgagtcgata cttcggcgat caccgcttcc ctcatactct
     4441 tcctttttca ttattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4501 ttgaatgtat ttagaaaaat aaacaaatag ctagctcact cggtcgctac gctccgggcg
     4561 tgagactgcg gcgggcgctg cggacacata caaagttacc cacagattcc gtggataagc
     4621 aggggactaa catgtgaggc aaaacagcag ggccgcgccg gtggcgtttt tccataggct
     4681 ccgccctcct gccagagttc acataaacag acgcttttcc ggtgcatctg tgggagccgt
     4741 gaggctcaac catgaatctg acagtacggg cgaaacccga caggacttaa agatccccac
     4801 cgtttccggc gggtcgctcc ctcttgcgct ctcctgttcc gaccctgccg tttaccggat
     4861 acctgttccg cctttctccc ttacgggaag tgtggcgctt tctcatagct cacacactgg
     4921 tatctcggct cggtgtaggt cgttcgctcc aagctgggct gtaagcaaga actccccgtt
     4981 cagcccgact gctgcgcctt atccggtaac tgttcacttg agtccaaccc ggaaaagcac
     5041 ggtaaaacgc cactggcagc agccattggt aactgggagt tcgcagagga tttgtttagc
     5101 taaacacgcg gttgctcttg aagtgtgcgc caaagtccgg ctacactgga aggacagatt
     5161 tggttgctgt gctctgcgaa agccagttac cacggttaag cagttcccca actgacttaa
     5221 ccttcgatca aaccacctcc ccaggtggtt ttttcgaagg ttaagtcagt tggggaactg
     5281 cttaaccttc gatcaaacca cctccccagg tggttttttc gtttacaggg caaaagatta
     5341 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctactgaa ccgctcttga
     5401 tttcagtgca atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt
     5461 tgtaattctc atgttagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     5521 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
     5581 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5641 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca
     5701 ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca
     5761 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg
     5821 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg tccgcaccaa
     5881 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
     5941 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
     6001 a
//