LOCUS       NM4053                  2766 bp    DNA        circular     30-JUL-2023
DEFINITION  .
ACCESSION   
VERSION     
SOURCE      .
  ORGANISM  .
COMMENT     Golden Gate reaction:
COMMENT     DR274 5arm-BsaI [3467] from 1714 (BsaI) to 1677 (BsaI) 
COMMENT     cxti10882 left arm PCR from 8 (BsaI) to 888 (BsaI) 
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     CDS             complement(686..1501)
                     /locus_tag="KanR2"
                     /label="KanR2"
                     /ApEinfo_label="KanR2"
                     /ApEinfo_fwdcolor="#66ccff"
                     /ApEinfo_revcolor="#66ccff"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    complement(2521..2522)
                     /locus_tag="Mos "
                     /label="Mos "
                     /ApEinfo_label="Mos "
                     /ApEinfo_fwdcolor="#fefb00"
                     /ApEinfo_revcolor="#fefb00"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    2482..2504
                     /locus_tag="sgrNS site 1"
                     /label="sgrNS site 1"
                     /ApEinfo_label="sgrNS site 1"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     rep_origin      complement(36..623)
                     /locus_tag="Ori"
                     /label="Ori"
                     /ApEinfo_label="Ori"
                     /ApEinfo_fwdcolor="#808080"
                     /ApEinfo_revcolor="#808080"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     1580..1599
                     /locus_tag="NMoligo 5008"
                     /label="NMoligo 5008"
                     /ApEinfo_label="NMoligo 5008"
                     /ApEinfo_fwdcolor="#808000"
                     /ApEinfo_revcolor="#800040"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_binding    1672..1678
                     /locus_tag="SapI >"
                     /label="SapI >"
                     /ApEinfo_label="SapI >"
                     /ApEinfo_fwdcolor="#ff0000"
                     /ApEinfo_revcolor="#ff0000"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    2544..2559
                     /locus_tag="sgRNA site 2"
                     /label="sgRNA site 2"
                     /ApEinfo_label="sgRNA site 2"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    1683..2559
                     /locus_tag="arm1"
                     /label="arm1"
                     /ApEinfo_label="arm1"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    2556..2559
                     /locus_tag="RIGHT ARM "
                     /label="RIGHT ARM "
                     /ApEinfo_label="RIGHT ARM "
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     2029..2048
                     /locus_tag="NMoligo 3890"
                     /label="NMoligo 3890"
                     /ApEinfo_label="NMoligo 3890"
                     /ApEinfo_fwdcolor="#808000"
                     /ApEinfo_revcolor="#800040"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     2541..2559
                     /locus_tag="NMoligo 4029"
                     /label="NMoligo 4029"
                     /ApEinfo_label="NMoligo 4029"
                     /ApEinfo_fwdcolor="#808000"
                     /ApEinfo_revcolor="#800040"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     complement(2745..2762)
                     /locus_tag="M13-fwd"
                     /label="M13-fwd"
                     /ApEinfo_label="M13-fwd"
                     /ApEinfo_fwdcolor="#808000"
                     /ApEinfo_revcolor="#800040"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    2564..2570
                     /locus_tag="< SapI "
                     /label="< SapI "
                     /ApEinfo_label="< SapI "
                     /ApEinfo_fwdcolor="#ff0000"
                     /ApEinfo_revcolor="#ff0000"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     enzyme          2564..2570
                     /note="GCTCTTC(1/4)"
                     /locus_tag="SapI"
                     /label="SapI"
                     /ApEinfo_label="SapI"
                     /ApEinfo_fwdcolor="#00DF87"
                     /ApEinfo_revcolor="#00DF87"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
ORIGIN
        1 TGGGCCTTTC TTCGGTAGAA AATCAAAGGA TCTTCttgag atcctttttt tctgcgcgta
       61 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
      121 gagctaccaa ctctttttcc gaggtaactg gcttcagcag agcgcagata ccaaatactg
      181 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
      241 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
      301 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
      361 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
      421 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
      481 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
      541 tttatagtcc tgtcgggttt cgccacctct gacttgagca tcgatttttg tgatgctcgt
      601 caggggggcg gagcctatgg aaaAACGCCA GCAACGCAGA AAGGCCCACC CGAAGGTGAG
      661 CCAGGTGATT ACATTTAGGT CCTCATTAGA AAAACTCATC GAGCATCAAG TGAAACTGCA
      721 ATTTATTCAT ATCAGGATTA TCAATACCAT ATTTTTGAAA AAGCCGTTTC TGTAATGAAG
      781 GAGAAAACTC ACCGAGGCAG TTCCATAGGA TGGCAAGATC CTGGTATCGG TCTGCGATTC
      841 CGACTCGTCC AACATCAATA CAACCTATTA ATTTCCCCTC GTCAAAAATA AGGTTATCAA
      901 GTGAGAAATC ACCATGAGTG ACGACTGAAT CCGGTGAGAA TGGCAAGAGT TTATGCATTT
      961 CTTTCCAGAC TTGTTCAACA GGCCAGCCAT TACGCTCGTC ATCAAAATCA CTCGCACCAA
     1021 CCAAACCGTT ATTCATTCGT GATTGCGCCT GAGCGAGACG AAATACGCGA TCGCCGTTAA
     1081 AAGGACAATT ACAAACAGGA ATCGAATGCA ACCGGCGCAG GAACACTGCC AGCGCATCAA
     1141 CAATATTTTC ACCTGAATCA GGATATTCTT CTAATACCTG GAATGCTGTT TTCCCTGGGA
     1201 TCGCAGTGGT GAGTAACCAT GCATCATCAG GAGTACGGAT AAAATGCTTG ATGGTCGGAA
     1261 GAGGCATAAA TTCCGTCAGC CAGTTTAGCC TGACCATCTC ATCTGTAACA TCATTGGCAA
     1321 CGCTACCTTT GCCATGTTTC AGAAACAACT CTGGCGCATC GGGCTTCCCA TACAATCGAT
     1381 AGATTGTCGC ACCTGATTGC CCGACATTAT CGCGAGCCCA TTTATACCCA TATAAATCAG
     1441 CATCCATGTT GGAATTTAAT CGCGGCTTCA AGCAAGACGT TTCCCGTTGA ATATGGCTCA
     1501 TTTTAGCTTC CTTAGCTCCT GAAAATCTCG ATAACTCAAA AAATACGCCC GGTAGTGATC
     1561 TTATTTCATT ATGGTGAAAg ttggaacctc ttacgtgccG ATCAAGTCAA AAGCCTCCGG
     1621 TCGGAGGCTT TTGACTTTCT GCTATGGAGG TCAGGTATGA TTTttGAATT CGCTCTTCgT
     1681 GGtttcaacg gtggaagaag gggcaaattt tataccaaaa cgtctccgtt tatgacccac
     1741 cccctatgca aaataaacgg ctctgggaga aatagtacag aaaacgcgga gcctgcttga
     1801 ttccttctcc gaggcgacaa aaaacgcatc attcgtattg taagatgcat gtgtggctgg
     1861 cctcttttga attttactgg ttttttatat ttctactaat atttcgaaaa attttgtaag
     1921 taaatcattg attacgttta caaaaattta gaaacatttc aaaatgatta cttctaaatt
     1981 aattatattt acaggttagt tcgagttact tatgaccttc caatccgcca tatccgccaa
     2041 ggacgctcca attaattttt tagaattatt ttttaacaca tgtattattt tttaacaata
     2101 aattattcat gaaatcatca aatgtatatg ttctactata atatttaggt caaactatag
     2161 aaaagaaatc cttgtttttg gaacaaagga gttcagatcc tgtgcatttg atcttggatc
     2221 cacatctttg gtccatggga aagcttggca ctgatcaaga aggattcttc aaatatctaa
     2281 aattgccggc tgcccgtaga attccatcag ttttccagag ttttagtgaa aatttgcatc
     2341 ccgttagaaa caatctacac aacggtaaaa aggacccatt atttttaccg taccgttttc
     2401 ctttatggcc ctctatgcaa aaaattatag cgtggagggg cagtacagaa taataaagtg
     2461 tgagaaaaaa aactgctctg cgagaaatag tacagcaaac gcggagcatg cttgattcta
     2521 taattcaaaa acgcagaact cgtactgttg gatgcctgtG CGAGAAGAGC AAGCTTGGAT
     2581 CGACGAGAGC AGCGCGACTG GATCTGTCGC CCGTCTCAAA CGCAACCCTC CGGCGGTCGC
     2641 ATATCATTCA GGACGAGCCT CAGACTCCAG CGTAACTGGA CTGCAATCAA CTCACTGGCT
     2701 CACCTTCCGG TCCACGATCA GCTAGAATCA AGCTGACTAG ATAAACTGGC CGTCGTTTTA
     2761 CACGGG     
//
