LOCUS       pENTR-BS-AtMIR390a-A18G 4080 bp ds-DNA     circular     31-JUL-2025
DEFINITION  .
KEYWORDS    "marker:M13-R, KanR, CmR"
FEATURES             Location/Qualifiers
     terminator      complement(268..295)
                     /label="rrnB T2 terminator"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /note="label: rrnB T2"
     terminator      complement(427..470)
                     /label="rrnB T1 terminator"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /note="label: rrnB T1"
     primer_bind     536..553
                     /label="M13-F"
                     /ApEinfo_revcolor="#7f007f"
                     /ApEinfo_fwdcolor="#7f007f"
                     /note="label: M13-F"
     primer_bind     536..553
                     /label="M13-fwd"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
                     /note="label: M13-fwd"
     misc_recomb     569..668
                     /label="AttL1"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /note="label: AttL1"
     misc_recomb     570..665
                     /label="AttL1"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
                     /note="label: AttL1"
     misc_feature    645..665
                     /label="Gateway recomb1"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /note="label: Gateway recomb1"
     misc_recomb     645..665
                     /label="attB1"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
                     /note="label: attB1"
     ncRNA           689..702
                     /label="AtMIR390a 5' region"
                     /ApEinfo_revcolor="#0000ff"
                     /ApEinfo_fwdcolor="#0000ff"
                     /note="label: AtMIR390a 5' region"
     misc_feature    706..706
                     /label="a18g"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     promoter        730..735
                     /label="lac UV5 promoter"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
                     /note="note: E. coli lac promoter with an ""up"" mutation"
     promoter        736..753
                     /label="lac UV5 promoter"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
                     /note="note: E. coli lac promoter with an ""up"" mutation"
     promoter        754..760
                     /label="lac UV5 promoter"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
                     /note="note: E. coli lac promoter with an ""up"" mutation"
     CDS             814..1473
                     /label="CmR"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
                     /note="codon_start: 1 translation: MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFLKTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSSLWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANMDNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA* gene: cat product: chloramphenicol acetyltransferase transl_table: 1 note: confers resistance to chloramphenicol"
     CDS             1815..2120
                     /label="ccdB"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
                     /note="codon_start: 1 translation: MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKVSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI* gene: ccdB product: CcdB, a bacterial toxin that poisons DNA gyrase transl_table: 1 note: Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains."
     ncRNA           2168..2184
                     /label="AtMIR390a 3' region"
                     /ApEinfo_revcolor="#0000ff"
                     /ApEinfo_fwdcolor="#0000ff"
                     /note="label: AtMIR390a 3' region"
     misc_recomb     complement(2205..2304)
                     /label="AttL2"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /note="label: AttL2"
     misc_recomb     complement(2208..2228)
                     /label="attB2"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
                     /note="label: attB2"
     misc_feature    complement(2208..2228)
                     /label="Gateway Recomb2"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /note="label: Gateway Recomb2"
     misc_recomb     complement(2208..2303)
                     /label="AttL2"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
                     /note="label: AttL2"
     promoter        complement(2321..2340)
                     /label="T7 promoter"
                     /ApEinfo_revcolor="#008b8b"
                     /ApEinfo_fwdcolor="#008b8b"
                     /note="label: T7"
     primer_bind     complement(2321..2342)
                     /label="T7"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
                     /note="label: T7"
     primer_bind     complement(2341..2361)
                     /label="M13-R"
                     /ApEinfo_revcolor="#7f007f"
                     /ApEinfo_fwdcolor="#7f007f"
                     /note="label: M13-R"
     primer_bind     complement(2341..2361)
                     /label="M13-rev"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
                     /note="label: M13-rev"
     CDS             2474..3280
                     /label="Kanamycin resistance gene"
                     /ApEinfo_revcolor="#996633"
                     /ApEinfo_fwdcolor="#996633"
                     /note="label: Kanamycin resistance gene"
     CDS             2474..3280
                     /label="KanR"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /note="label: KanR"
     rep_origin      3408..4036
                     /label="ColE1 origin"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
                     /note="label: ColE1 origin"
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 CCTTCACCAG TAGAGAAGAA TCTGTgAGAG ACCATTAGGC
      721 ACCCCAGGCT TTACACTTTA TGCTTCCGGC TCGTATAATG TGTGGATTTT GAGTTAGGAG
      781 CCGTCGAGAT TTTCAGGAGC TAAGGAAGCT AAAatggaga aaaaaatcac tggatatacc
      841 accgttgata tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct
      901 caatgtacct ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag
      961 aaaaataagc acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct
     1021 catccggagt tccgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac
     1081 ccttgttaca ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac
     1141 cacgacgatt tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa
     1201 aacctggcct atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc
     1261 tgggtgagtt tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc
     1321 gttttcacca tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt
     1381 caggttcatc atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa
     1441 cagtactgcg atgagtggca gggcggggcg taaACGCGTG GAGCCGGCTT ACTAAAAGCC
     1501 AGATAACAGT ATGCGTATTT GCGCGCTGAT TTTTGCGGTA TAAGAATATA TACTGATATG
     1561 TATACCCGAA GTATGTCAAA AAGAGGTATG CTATGAAGCA GCGTATTACA GTGACAGTTG
     1621 ACAGCGACAG CTATCAGTTG CTCAAGGCAT ATATGATGTC AATATCTCCG GTCTGGTAAG
     1681 CACAACCATG CAGAATGAAG CCCGTCGTCT GCGTGCCGAA CGCTGGAAAG CGGAAAATCA
     1741 GGAAGGGATG GCTGAGGTCG CCCGGTTTAT TGAAATGAAC GGCTCTTTTG CTGACGAGAA
     1801 CAGGGGCTGG TGAAATGCAG TTTAAGGTTT ACACCTATAA AAGAGAGAGC CGTTATCGTC
     1861 TGTTTGTGGA TGTACAGAGT GATATTATTG ACACGCCCGG CCGACGGATG GTGATCCCCC
     1921 TGGCCAGTGC ACGTCTGCTG TCAGATAAAG TCTCCCGTGA ACTTTACCCG GTGGTGCATA
     1981 TCGGGGATGA AAGCTGGCGC ATGATGACCA CCGATATGGC CAGTGTGCCG GTTTCCGTTA
     2041 TCGGGGAAGA AGTGGCTGAT CTCAGCCACC GCGAAAATGA CATCAAAAAC GCCATTAACC
     2101 TGATGTTCTG GGGAATATAA ATGTCAGGCT CCCTTATACA CAGCCAGTCT GCACCTCGAC
     2161 ggtctcAcat tggctcttct tactCATTAA GGGTGGGCGC GCCGACCCAG CTTTCTTGTA
     2221 CAAAGTTGGC ATTATAAGAA AGCATTGCTT ATCAATTTGT TGCAACGAAC AGGTCACTAT
     2281 CAGTCAAAAT AAAATCATTA TTTGCCATCC AGCTGATATC CCCTATAGTG AGTCGTATTA
     2341 CATGGTCATA GCTGTTTCCT GGCAGCTCTG GCCCGTGTCT CAAAATCTCT GATGTTACAT
     2401 TGCACAAGAT AAAAATATAT CATCATGAAC AATAAAACTG TCTGCTTACA TAAACAGTAA
     2461 TACAAGGGGT GTTatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa
     2521 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     2581 gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     2641 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     2701 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     2761 cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     2821 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     2881 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     2941 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     3001 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     3061 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     3121 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     3181 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     3241 gaataaattg cagtttcatt tgatgctcga tgagtttttc TAATCAGAAT TGGTTAATTG
     3301 GTTGTAACAC TGGCAGAGCA TTACGCTGAC TTGACGGGAC GGCGCAAGCT CATGACCAAA
     3361 ATCCCTTAAC GTGAGTTACG CGTCGTTCCA CTGAGCGTCA GACCCCGTAG AAAAGATCAA
     3421 AGGATCTTCT TGAGATCCTT TTTTTCTGCG CGTAATCTGC TGCTTGCAAA CAAAAAAACC
     3481 ACCGCTACCA GCGGTGGTTT GTTTGCCGGA TCAAGAGCTA CCAACTCTTT TTCCGAAGGT
     3541 AACTGGCTTC AGCAGAGCGC AGATACCAAA TACTGTCCTT CTAGTGTAGC CGTAGTTAGG
     3601 CCACCACTTC AAGAACTCTG TAGCACCGCC TACATACCTC GCTCTGCTAA TCCTGTTACC
     3661 AGTGGCTGCT GCCAGTGGCG ATAAGTCGTG TCTTACCGGG TTGGACTCAA GACGATAGTT
     3721 ACCGGATAAG GCGCAGCGGT CGGGCTGAAC GGGGGGTTCG TGCACACAGC CCAGCTTGGA
     3781 GCGAACGACC TACACCGAAC TGAGATACCT ACAGCGTGAG CATTGAGAAA GCGCCACGCT
     3841 TCCCGAAGGG AGAAAGGCGG ACAGGTATCC GGTAAGCGGC AGGGTCGGAA CAGGAGAGCG
     3901 CACGAGGGAG CTTCCAGGGG GAAACGCCTG GTATCTTTAT AGTCCTGTCG GGTTTCGCCA
     3961 CCTCTGACTT GAGCGTCGAT TTTTGTGATG CTCGTCAGGG GGGCGGAGCC TATGGAAAAA
     4021 CGCCAGCAAC GCGGCCTTTT TACGGTTCCT GGCCTTTTGC TGGCCTTTTG CTCACATGTT
//