LOCUS       DR274_FP_nls_mNG        2891 bp    DNA        circular     02-AUG-2023
DEFINITION  .
ACCESSION   
VERSION     
SOURCE      .
  ORGANISM  .
COMMENT     Ligation reaction:
COMMENT     DR274 FP tomm-20 [4259] from 2663 (SapI) to 1678 (SapI)
COMMENT     PCR DR274 NT nls mNG 7070 7071 from 12 (SapI) to 1016 (SapI)
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"
     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"
     misc_feature    1680..1682
                     /locus_tag="pMYO-2"
                     /label="pMYO-2"
                     /ApEinfo_label="pMYO-2"
                     /ApEinfo_fwdcolor="#85dae9"
                     /ApEinfo_revcolor="#85dae9"
                     /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_structure  1683..1688
                     /locus_tag="BamHI"
                     /label="BamHI"
                     /ApEinfo_label="BamHI"
                     /ApEinfo_fwdcolor="#4c4c4c"
                     /ApEinfo_revcolor="#4c4c4c"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    2457..2507
                     /locus_tag="Synthetic Intron 4"
                     /label="Synthetic Intron 4"
                     /ApEinfo_label="Synthetic Intron 4"
                     /ApEinfo_fwdcolor="#ff80c0"
                     /ApEinfo_revcolor="#ff80c0"
                     /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"
     primer_bind     complement(2671..2684)
                     /note=") PCR_conditions {primer
                     sequence:ccttGCTCTTCcCTTGGCGAGCTTCTTGG}"
                     /locus_tag="7071"
                     /label="7071"
                     /ApEinfo_label="7071"
                     /ApEinfo_fwdcolor="#408000"
                     /ApEinfo_revcolor="#800080"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    join(1713..1818,1870..1998,2050..2180,2232..2456,
                     2508..2621)
                     /locus_tag="NeonGreen_Cel"
                     /label="NeonGreen_Cel"
                     /ApEinfo_label="NeonGreen_Cel"
                     /ApEinfo_fwdcolor="#00ff00"
                     /ApEinfo_revcolor="#00ff00"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     sig_peptide     1689..1711
                     /locus_tag="SV40"
                     /label="SV40"
                     /ApEinfo_label="SV40"
                     /ApEinfo_fwdcolor="#ff0080"
                     /ApEinfo_revcolor="#ff0080"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     1680..1698
                     /note=") PCR_conditions {primer
                     sequence:TTGTGCTCTTCaATGGGATCCCCAAAGAAGA}"
                     /locus_tag="7070"
                     /label="7070"
                     /ApEinfo_label="7070"
                     /ApEinfo_fwdcolor="#408000"
                     /ApEinfo_revcolor="#800080"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    2181..2231
                     /locus_tag="Synthetic Intron 3"
                     /label="Synthetic Intron 3"
                     /ApEinfo_label="Synthetic Intron 3"
                     /ApEinfo_fwdcolor="#ff80c0"
                     /ApEinfo_revcolor="#ff80c0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    1819..1869
                     /locus_tag="Synthetic Intron 1"
                     /label="Synthetic Intron 1"
                     /ApEinfo_label="Synthetic Intron 1"
                     /ApEinfo_fwdcolor="#ff80c0"
                     /ApEinfo_revcolor="#ff80c0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    1999..2049
                     /locus_tag="Synthetic Intron 2"
                     /label="Synthetic Intron 2"
                     /ApEinfo_label="Synthetic Intron 2"
                     /ApEinfo_fwdcolor="#ff80c0"
                     /ApEinfo_revcolor="#ff80c0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     transit_peptide 2628..2684
                     /locus_tag="egl-13 NLS"
                     /label="egl-13 NLS"
                     /ApEinfo_label="egl-13 NLS"
                     /ApEinfo_fwdcolor="#ff0080"
                     /ApEinfo_revcolor="#ff0080"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     complement(2870..2887)
                     /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    2689..2695
                     /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"
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 CGCTCTTCgA
     1681 TGGGATCCCC AAAGAAGAAG CGTAAGGTGT CgATGGTGTC GAAGGGAGAA GAGGATAACA
     1741 TGGCTTCACT CCCAGCTACA CACGAACTCC ACATCTTCGG ATCGATCAAC GGAGTGGATT
     1801 TCGATATGGT CGGACAAGgt aagtttaaac atatatatac taactaaccc tgattattta
     1861 aattttcagG AACTGGAAAC CCAAACGATG GATACGAGGA ACTCAACCTC AAGTCGACAA
     1921 AGGGAGATCT GCAATTCTCG CCATGGATTC TCGTGCCACA CATCGGATAC GGATTCCACC
     1981 AATACCTCCC ATACCCAGgt aagtttaaac tgagttctac taactaacga gtaatattta
     2041 aattttcagA TGGAATGTCA CCATTCCAAG CTGCCATGGT GGATGGATCG GGATACCAAG
     2101 TTCACCGAAC AATGCAATTC GAGGATGGAG CCTCGCTCAC AGTGAACTAC CGATACACAT
     2161 ACGAGGGATC GCACATCAAG gtaagtttaa acagttcggt actaactaac catacatatt
     2221 taaattttca gGGAGAGGCT CAAGTTAAGG GAACAGGATT CCCAGCTGAT GGACCAGTGA
     2281 TGACAAACTC ACTCACAGCT GCTGATTGGT GCCGATCGAA AAAGACATAC CCAAATGATA
     2341 AGACAATCAT CTCGACATTC AAGTGGTCGT ACACTACTGG AAACGGAAAG CGATACCGAT
     2401 CGACAGCCCG AACAACATAC ACATTCGCTA AGCCAATGGC CGCCAACTAC CTCAAGgtaa
     2461 gtttaaacat gattttacta actaactaat ctgatttaaa ttttcagAAT CAACCAATGT
     2521 ACGTGTTCCG AAAGACAGAA CTCAAGCACT CAAAGACAGA GCTGAACTTC AAAGAGTGGC
     2581 AAAAGGCCTT CACAGATGTG ATGGGAATGG ATGAACTCTA CAAGGCCTCC CGTCGTCGTA
     2641 AGGCCAACCC AACCAAGCTC TCCGAGAACG CCAAGAAGCT CGCCAAGgGA AGAGCAAGCT
     2701 TGGATCGACG AGAGCAGCGC GACTGGATCT GTCGCCCGTC TCAAACGCAA CCCTCCGGCG
     2761 GTCGCATATC ATTCAGGACG AGCCTCAGAC TCCAGCGTAA CTGGACTGCA ATCAACTCAC
     2821 TGGCTCACCT TCCGGTCCAC GATCAGCTAG AATCAAGCTG ACTAGATAAA CTGGCCGTCG
     2881 TTTTACACGG G    
//
