LOCUS       pENTR-SlymiR482bTS-Bc_A 4086 bp ds-DNA     circular     31-MAR-2023
DEFINITION  .
FEATURES             Location/Qualifiers
     terminator      complement(268..295)
                     /label="rrnB T2"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     primer          complement(318..336)
                     /label="AC-588"
                     /note="sequence: TTGGAATGATGCCCACCATAA"
                     /ApEinfo_revcolor="#f7977a"
                     /ApEinfo_fwdcolor="#f7977a"
     primer          complement(318..336)
                     /label="AC-471"
                     /note="sequence: AATGTTATGGTGGGAATCATTCCAAAAAAAATAGATAACGAATGTGATCATCATTTGGAATGATGCCCACCATAA"
                     /ApEinfo_revcolor="#f7977a"
                     /ApEinfo_fwdcolor="#f7977a"
     terminator      complement(427..470)
                     /label="rrnB T1"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     primer_bind     536..553
                     /label="M13-fwd"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer_bind     536..553
                     /label="M13-F"
                     /ApEinfo_revcolor="#7f007f"
                     /ApEinfo_fwdcolor="#7f007f"
     primer          536..553
                     /label="AC-33"
                     /note="sequence: tgtaaaacgacggccagt"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     misc_recomb     569..668
                     /label="AttL1"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_recomb     570..665
                     /label="AttL1"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_feature    645..665
                     /label="Gateway recomb1"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_recomb     645..665
                     /label="attB1"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     primer          671..692
                     /label="AC-335"
                     /note="sequence: CACCAGTAGAGAAGAATCTGTA"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          complement(674..688)
                     /label="AC-23"
                     /note="sequence: CATGGTGAAGGGGGCGGC"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer          complement(674..688)
                     /label="D2475"
                     /note="sequence: CATGGTGAAGGGGGCGGC"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     ncRNA           689..692
                     /label="AtMIR390a 5' region"
                     /ApEinfo_revcolor="#0000ff"
                     /ApEinfo_fwdcolor="#0000ff"
     misc_feature    693..714
                     /label="SlymiR482bTS"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     misc_feature    715..725
                     /label="AtTAS1c spacer"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    727..732
                     /label="BsaI site"
                     /ApEinfo_revcolor="#ff0000"
                     /ApEinfo_fwdcolor="#ff0000"
     misc_feature    833..1492
                     /label="Chloramphenicol resistance gene"
                     /ApEinfo_revcolor="#fb02fe"
                     /ApEinfo_fwdcolor="#fb02fe"
     ncRNA           1147..1147
                     /label="AtTAS1c 3' region [5']"
                     /ApEinfo_revcolor="#0000ff"
                     /ApEinfo_fwdcolor="#0000ff"
     misc_feature    1834..2139
                     /label="ccdB"
                     /ApEinfo_revcolor="#fb02fe"
                     /ApEinfo_fwdcolor="#fb02fe"
     misc_feature    2180..2185
                     /label="Inverted BsaI site"
                     /ApEinfo_revcolor="#ff0000"
                     /ApEinfo_fwdcolor="#ff0000"
     primer          complement(2186..2203)
                     /label="AC-336"
                     /note="sequence: AGTAAGAAGAGCCAATGT"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     ncRNA           2191..2194
                     /label="AtMIR390a 3' region"
                     /ApEinfo_revcolor="#0000ff"
                     /ApEinfo_fwdcolor="#0000ff"
     misc_recomb     complement(2211..2310)
                     /label="AttL2"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    complement(2214..2234)
                     /label="Gateway Recomb2"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_recomb     complement(2214..2234)
                     /label="attB2"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_recomb     complement(2214..2309)
                     /label="AttL2"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     promoter        complement(2327..2346)
                     /label="T7"
                     /ApEinfo_revcolor="#008b8b"
                     /ApEinfo_fwdcolor="#008b8b"
     primer_bind     complement(2327..2348)
                     /label="T7"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer_bind     complement(2347..2367)
                     /label="M13-rev"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer_bind     complement(2347..2367)
                     /label="M13-R"
                     /ApEinfo_revcolor="#7f007f"
                     /ApEinfo_fwdcolor="#7f007f"
     primer          complement(2347..2367)
                     /label="AC-17"
                     /note="sequence: CAGGAAACAGCTATGACCATG"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     CDS             2480..3286
                     /label="Kanamycin resistance gene"
                     /ApEinfo_revcolor="#996633"
                     /ApEinfo_fwdcolor="#996633"
     CDS             2480..3286
                     /label="KanR"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     primer          complement(2787..2807)
                     /label="AC-331"
                     /note="sequence: CACCGTGTAATACTTGTAACGCTG"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     rep_origin      3414..4042
                     /label="ColE1 origin"
                     /ApEinfo_revcolor="#4682b4"
                     /ApEinfo_fwdcolor="#4682b4"
ORIGIN
        1 CTTTCCTGCG TTATCCCCTG ATTCTGTGGA TAACCGTATT ACCGCCTTTG AGTGAGCTGA
       61 TACCGCTCGC CGCAGCCGAA CGACCGAGCG CAGCGAGTCA GTGAGCGAGG AAGCGGAAGA
      121 GCGCCCAATA CGCAAACCGC CTCTCCCCGC GCGTTGGCCG ATTCATTAAT GCAGCTGGCA
      181 CGACAGGTTT CCCGACTGGA AAGCGGGCAG TGAGCGCAAC GCAATTAATA CGCGTACCGC
      241 TAGCCAGGAA GAGTTTGTAG AAACGCAAAA AGGCCATCCG TCAGGATGGC CTTCTGCTTA
      301 GTTTGATGCC TGGCAGTTTA TGGCGGGCGT CCTGCCCGCC ACCCTCCGGG CCGTTGCTTC
      361 ACAACGTTCA AATCCGCTCC CGGCGGATTT GTCCTACTCA GGAGAGCGTT CACCGACAAA
      421 CAACAGATAA AACGAAAGGC CCAGTCTTCC GACTGAGCCT TTCGTTTTAT TTGATGCCTG
      481 GCAGTTCCCT ACTCTCGCGT TAACGCTAGC ATGGATGTTT TCCCAGTCAC GACGTTGTAA
      541 AACGACGGCC AGTCTTAAGC TCGGGCCCCA AATAATGATT TTATTTTGAC TGATAGTGAC
      601 CTGTTCGTTG CAACAAATTG ATGAGCAATG CTTTTTTATA ATGCCAACTT TGTACAAAAA
      661 AGCAGGCTCC GCGGCCGCCC CCTTCACCTG TAGGCATGGG CGGTGTAGGC AAGATAGACC
      721 ATTTAAGAGA CCATTAGGCA CCCCAGGCTT TACACTTTAT GCTTCCGGCT CGTATAATGT
      781 GTGGATTTTG AGTTAGGAGC CGTCGAGATT TTCAGGAGCT AAGGAAGCTA AAatggagaa
      841 aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga
      901 ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc
      961 ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct
     1021 tgcccgcctg atgaatgctc atccggagtt ccgtatggca atgaaagacg gtgagctggt
     1081 gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc
     1141 atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga
     1201 tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt
     1261 tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat
     1321 ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt
     1381 gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag
     1441 aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt aaACGCGTGG
     1501 AGCCGGCTTA CTAAAAGCCA GATAACAGTA TGCGTATTTG CGCGCTGATT TTTGCGGTAT
     1561 AAGAATATAT ACTGATATGT ATACCCGAAG TATGTCAAAA AGAGGTATGC TATGAAGCAG
     1621 CGTATTACAG TGACAGTTGA CAGCGACAGC TATCAGTTGC TCAAGGCATA TATGATGTCA
     1681 ATATCTCCGG TCTGGTAAGC ACAACCATGC AGAATGAAGC CCGTCGTCTG CGTGCCGAAC
     1741 GCTGGAAAGC GGAAAATCAG GAAGGGATGG CTGAGGTCGC CCGGTTTATT GAAATGAACG
     1801 GCTCTTTTGC TGACGAGAAC AGGGGCTGGT GAAATGCAGT TTAAGGTTTA CACCTATAAA
     1861 AGAGAGAGCC GTTATCGTCT GTTTGTGGAT GTACAGAGTG ATATTATTGA CACGCCCGGC
     1921 CGACGGATGG TGATCCCCCT GGCCAGTGCA CGTCTGCTGT CAGATAAAGT CTCCCGTGAA
     1981 CTTTACCCGG TGGTGCATAT CGGGGATGAA AGCTGGCGCA TGATGACCAC CGATATGGCC
     2041 AGTGTGCCGG TTTCCGTTAT CGGGGAAGAA GTGGCTGATC TCAGCCACCG CGAAAATGAC
     2101 ATCAAAAACG CCATTAACCT GATGTTCTGG GGAATATAAA TGTCAGGCTC CCTTATACAC
     2161 AGCCAGTCTG CACCTCGACg gtctcATCGG cattAAGGGT GGGCGCGCCG ACCCAGCTTT
     2221 CTTGTACAAA GTTGGCATTA TAAGAAAGCA TTGCTTATCA ATTTGTTGCA ACGAACAGGT
     2281 CACTATCAGT CAAAATAAAA TCATTATTTG CCATCCAGCT GATATCCCCT ATAGTGAGTC
     2341 GTATTACATG GTCATAGCTG TTTCCTGGCA GCTCTGGCCC GTGTCTCAAA ATCTCTGATG
     2401 TTACATTGCA CAAGATAAAA ATATATCATC ATGAACAATA AAACTGTCTG CTTACATAAA
     2461 CAGTAATACA AGGGGTGTTa tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     2521 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     2581 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     2641 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     2701 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     2761 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     2821 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     2881 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     2941 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     3001 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     3061 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     3121 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     3181 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     3241 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttcTAAT CAGAATTGGT
     3301 TAATTGGTTG TAACACTGGC AGAGCATTAC GCTGACTTGA CGGGACGGCG CAAGCTCATG
     3361 ACCAAAATCC CTTAACGTGA GTTACGCGTC GTTCCACTGA GCGTCAGACC CCGTAGAAAA
     3421 GATCAAAGGA TCTTCTTGAG ATCCTTTTTT TCTGCGCGTA ATCTGCTGCT TGCAAACAAA
     3481 AAAACCACCG CTACCAGCGG TGGTTTGTTT GCCGGATCAA GAGCTACCAA CTCTTTTTCC
     3541 GAAGGTAACT GGCTTCAGCA GAGCGCAGAT ACCAAATACT GTCCTTCTAG TGTAGCCGTA
     3601 GTTAGGCCAC CACTTCAAGA ACTCTGTAGC ACCGCCTACA TACCTCGCTC TGCTAATCCT
     3661 GTTACCAGTG GCTGCTGCCA GTGGCGATAA GTCGTGTCTT ACCGGGTTGG ACTCAAGACG
     3721 ATAGTTACCG GATAAGGCGC AGCGGTCGGG CTGAACGGGG GGTTCGTGCA CACAGCCCAG
     3781 CTTGGAGCGA ACGACCTACA CCGAACTGAG ATACCTACAG CGTGAGCATT GAGAAAGCGC
     3841 CACGCTTCCC GAAGGGAGAA AGGCGGACAG GTATCCGGTA AGCGGCAGGG TCGGAACAGG
     3901 AGAGCGCACG AGGGAGCTTC CAGGGGGAAA CGCCTGGTAT CTTTATAGTC CTGTCGGGTT
     3961 TCGCCACCTC TGACTTGAGC GTCGATTTTT GTGATGCTCG TCAGGGGGGC GGAGCCTATG
     4021 GAAAAACGCC AGCAACGCGG CCTTTTTACG GTTCCTGGCC TTTTGCTGGC CTTTTGCTCA
     4081 CATGTT
//