LOCUS       pSEM389___mosTI_        4478 bp    DNA        circular     26-JAN-2022
DEFINITION  .
COMMENT     Unpublished reagents. Sonia el Mouridi and Christian
            Froekjaer-Jensen. Synthetic Genome Biology lab. KAUST.
COMMENT     www.wormbuilder.org
COMMENT     MOdular Safe harbor Transgene Insertion (mosTI).
COMMENT     Cloning vector for single-copy insertion using unc-119 selection.
COMMENT     MCS/Golden Gate (BsaI) cloning vector with insertion upstream of
            mScarlet(2xNLS) and tbb-2 3' UTR.
COMMENT     BsaI overhangs: tgcc - mcs - aaaa (use aaaa as consensus start site
            for mScarlet with aaaaATG)
COMMENT     
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     CDS             join(70..369,421..527)
                     /locus_tag="cbr-unc-119 (non-rescuing exons 3-4)"
                     /label="cbr-unc-119 (non-rescuing exons 3-4)"
                     /ApEinfo_label="cbr-unc-119 (non-rescuing exons 3-4)"
                     /ApEinfo_fwdcolor="#9437ff"
                     /ApEinfo_revcolor="#9437ff"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     1..17
                     /locus_tag="M13F"
                     /label="M13F"
                     /ApEinfo_label="M13F"
                     /ApEinfo_fwdcolor="#72fa78"
                     /ApEinfo_revcolor="#ff7d78"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     complement(2383..2399)
                     /locus_tag="M13R"
                     /label="M13R"
                     /ApEinfo_label="M13R"
                     /ApEinfo_fwdcolor="#72fa78"
                     /ApEinfo_revcolor="#ff7d78"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    3400..4365
                     /locus_tag="AmpR"
                     /label="AmpR"
                     /ApEinfo_label="AmpR"
                     /ApEinfo_fwdcolor="#929292"
                     /ApEinfo_revcolor="#929292"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    2571..3159
                     /locus_tag="Ori"
                     /label="Ori"
                     /ApEinfo_label="Ori"
                     /ApEinfo_fwdcolor="#d5d5d5"
                     /ApEinfo_revcolor="#d5d5d5"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     3'UTR           528..640
                     /locus_tag="eft-3 3'UTR"
                     /label="eft-3 3'UTR"
                     /ApEinfo_label="eft-3 3'UTR"
                     /ApEinfo_fwdcolor="#f76ca1"
                     /ApEinfo_revcolor="#d783ff"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    641..674
                     /locus_tag="loxP"
                     /label="loxP"
                     /ApEinfo_label="loxP"
                     /ApEinfo_fwdcolor="#ffd478"
                     /ApEinfo_revcolor="#fefc78"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    680..750
                     /locus_tag="MCS(w 2xBsaI)"
                     /label="MCS(w 2xBsaI)"
                     /ApEinfo_label="MCS(w 2xBsaI)"
                     /ApEinfo_fwdcolor="#28ad48"
                     /ApEinfo_revcolor="#28ad48"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     3'UTR           1698..2027
                     /locus_tag="tbb-2 3'UTR (short)"
                     /label="tbb-2 3'UTR (short)"
                     /ApEinfo_label="tbb-2 3'UTR (short)"
                     /ApEinfo_fwdcolor="#d4fb78"
                     /ApEinfo_revcolor="#d4fb78"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    2028..2326
                     /locus_tag="Right homology"
                     /label="Right homology"
                     /ApEinfo_label="Right homology"
                     /ApEinfo_fwdcolor="#ffd478"
                     /ApEinfo_revcolor="#ffd478"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     CDS             join(756..806,858..1062,1114..1275,1327..1700)
                     /locus_tag="ce-mScarlet (NLS)"
                     /label="ce-mScarlet (NLS)"
                     /ApEinfo_label="ce-mScarlet (NLS)"
                     /ApEinfo_fwdcolor="#d783ff"
                     /ApEinfo_revcolor="#d783ff"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     sig_peptide     759..779
                     /locus_tag="SV40 NLS"
                     /label="SV40 NLS"
                     /ApEinfo_label="SV40 NLS"
                     /ApEinfo_fwdcolor="#ff84ff"
                     /ApEinfo_revcolor="#ff84ff"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     sig_peptide     1626..1697
                     /locus_tag="egl-13 NLS"
                     /label="egl-13 NLS"
                     /ApEinfo_label="egl-13 NLS"
                     /ApEinfo_fwdcolor="#ff84ff"
                     /ApEinfo_revcolor="#ff84ff"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
ORIGIN
        1 gtaaaacgac ggccagtcat gcataatccg cacgcatctg gaataaggaa gtgccattcc
       61 gcctgacctA AGTGCTTTTC GAGATCGCCA AACCGGAGAA CGATCAGGAG AATGATGAGT
      121 CGCCACAGGA GTCGGCAAGA TACGTGCGTT ATAGATTTGC TCCAAACTTT TTGAAACTCA
      181 AAACGGTCGG AGCAACTGTG GAATTCAAAG TAGGAGACAT CCCAATCCAT CATTTCCGAA
      241 TGATCGAACG TCACTTCTTC AAAGATCGCC TTCTGAAGTG TTTTGACTTT GAATTCGGAT
      301 TCTGTATTCC GAATTCACGA AACAACTGTG AACATATCTA TGAGTTCCCT CAACTCTCTC
      361 AACAACTCAg tgagtcatta ttctaaaaag tacaattcaa aagactaatc tctttttcag
      421 TGGACGACAT GATCAACAAT CCAAACGAGA CTCGTTCTGA CAGCTTCTAT TTCGTCGATA
      481 ACAAACTCGT CATGCACAAC AAAGCCGACT ACTCATATGA TGCATAAatc ttcattgttg
      541 agtttatctt gttgattttt gaataaatta tcaactcttt actttttaat gggttatgaa
      601 ataaataaac attgaaaact gataaacaac gttcatctct ataacttcgt atagcataca
      661 ttatacgaag ttattgcctg agaccagcta gcaagcttgc atgcctgcag gtcgactcta
      721 gaggatcccc gggtaccgag ctcaggtctc taaaaATGCC CAAAAAGAAA CGAAAGGTCG
      781 TTTCTAAGGG CGAAGCCGTC ATCAAGgtaa gttttacgaa attctgaagc tcacaaaaca
      841 tggagtacat ttttcagGAG TTCATGCGGT TCAAGGTCCA CATGGAGGGT AGTATGAACG
      901 GACACGAGTT CGAGATAGAG GGAGAGGGAG AGGGACGGCC ATACGAGGGA ACCCAAACGG
      961 CCAAGCTCAA GGTCACCAAA GGGGGACCAC TCCCATTCTC CTGGGATATC CTCTCCCCAC
     1021 AGTTCATGTA CGGAAGTCGT GCCTTCACCA AGCACCCCGC AGgtaagttt ataatctatg
     1081 atcttaattt ggcaatggaa aaagtttttt cagATATACC AGATTACTAT AAGCAGTCCT
     1141 TCCCAGAGGG ATTCAAATGG GAGCGTGTCA TGAACTTCGA GGATGGAGGA GCCGTCACCG
     1201 TCACGCAAGA CACGTCCCTC GAGGATGGAA CCCTAATCTA CAAGGTAAAG CTTCGTGGAA
     1261 CCAACTTCCC CCCAGgtaag ttttattcca taatacaaat atatattgaa caataatgct
     1321 tttcagATGG ACCAGTCATG CAAAAGAAAA CTATGGGATG GGAAGCCTCG ACCGAACGTC
     1381 TCTACCCAGA GGACGGGGTC CTCAAAGGAG ATATCAAGAT GGCCCTAAGA CTCAAGGACG
     1441 GAGGACGGTA TCTTGCCGAT TTCAAGACCA CGTACAAGGC TAAGAAGCCA GTACAAATGC
     1501 CAGGGGCCTA CAACGTAGAC CGTAAACTCG ACATCACGTC CCACAACGAG GACTACACGG
     1561 TCGTCGAACA ATACGAACGT TCCGAAGGAC GTCACTCCAC CGGAGGGATG GACGAGCTCT
     1621 ACAAGTCCCG GCGTCGTAAG GCCAACCCAA CGAAGCTCTC CGAGAACGCC AAAAAGCTCG
     1681 CCAAAGAGGT CGAGAATTAA atgcaaaatc ctttcaagca ttcccttctt ctctatcact
     1741 cttctttctt tttgtcaaaa aattctctcg ctaatttatt tgctttttta atgttattat
     1801 tttatgactt tttatagtca ctgaaaagtt tgcatctgag tgaagtgaat gctatcaaaa
     1861 tgtgattctg tctgatgtac tttcacaatc tctcttcaat tccattttga agtgctttaa
     1921 acccgaaagg ttgagaaaaa tgcgagcgct caaatatttg tattgtgttc gttgagtgac
     1981 ccaacaaaaa gaggaaactt tattgtgccg ccaagaaaaa agtctcacag gaaacggaga
     2041 atctgtgcgg caatgtctta tcatttgaaa cgcgccgtat cgatgctgac atcatgcggc
     2101 tcccgtgtta gtcgagaaga tgggtacggg cctttattca tccaacactc tacggcttct
     2161 tcaagagcta gtagggcacc ctgtagttgg aaagggatta tttcgtatgg cgagcccata
     2221 ccgtctttct tgcggaagct taacacgata ggaagttgga tagtttcgaa cgatggttcc
     2281 atacggaacg ctgtctcaag gatgagtctg acggagtgta actcgaaggc taggtggagg
     2341 ctcagtgatg ataagtctgc gatggtggat gcatgtgtca tggtcatagc tgtttcctgt
     2401 gtgaaattgt tatccgctca gagggcacaa tcctattccg cgctatccga caatctccaa
     2461 gacattaggt ggagttcagt tcggcgtatg gcatatgtcg ctggaaagaa catgtgagca
     2521 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2581 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2641 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2701 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2761 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2821 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2881 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2941 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3001 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3061 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3121 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3181 acggggtctg acgctctatt caacaaagcc gccgtcccgt caagtcagcg taaatgggta
     3241 gggggcttca aatcgtcctc gtgataccaa ttcggagcct gcttttttgt acaaacttgt
     3301 tgataatggc aattcaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3361 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3421 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3481 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3541 agagccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3601 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3661 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3721 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3781 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3841 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3901 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3961 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4021 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4081 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4141 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4201 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4261 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4321 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4381 agtgccagat acctgaaaca aaacccatcg tacggccaag gaagtctcca ataactgtga
     4441 tccaccacaa gcgccagggt tttcccagtc acgacgtt  
//
