LOCUS       pSEM386___mosTI_        4403 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COMMENT     Unpublished reagents.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 for mosTI(unc-119). Transgene
            inserted upstream of gfp and tbb-2 3' UTR. BsaI overhangs: tgcc -
            mcs - aaaa (use aaaa as consensus start site for gfp 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(2308..2324)
                     /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    3325..4290
                     /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    2496..3084
                     /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"
     CDS             join(756..861,913..1184,1236..1425,1477..1625)
                     /locus_tag="ce-gfp"
                     /label="ce-gfp"
                     /ApEinfo_label="ce-gfp"
                     /ApEinfo_fwdcolor="#72fa78"
                     /ApEinfo_revcolor="#72fa78"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     3'UTR           1623..1952
                     /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    1953..2251
                     /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"
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 taaaaATGTC AAAAGGAGAA GAACTCTTTA
      781 CCGGAGTCGT CCCAATTCTC GTCGAGCTCG ACGGAGACGT AAACGGACAC AAATTCTCGG
      841 TTTCCGGAGA AGGAGAAGGA Ggtaagtttt acgaaattct gaagctcaca aaacatggag
      901 tacatttttc agATGCTACT TATGGAAAAC TCACCCTTAA ATTCATTTGC ACCACCGGAA
      961 AGCTCCCAGT GCCGTGGCCG ACCCTTGTGA CTACATTCTG CTACGGAGTT CAATGCTTTT
     1021 CCCGCTACCC AGACCACATG AAGCGTCACG ATTTCTTCAA ATCAGCCATG CCAGAAGGAT
     1081 ACGTCCAGGA GCGAACAATT TTCTTCAAGG ACGATGGAAA CTACAAGACT CGTGCTGAAG
     1141 TCAAGTTTGA AGGAGATACT CTCGTGAACC GCATTGAGCT CAAGgtaagt ttataatcta
     1201 tgatcttaat ttggcaatgg aaaaagtttt ttcagGGAAT CGACTTCAAA GAAGATGGAA
     1261 ACATTCTTGG ACACAAGCTC GAATACAACT ATAACTCGCA CAACGTGTAT ATCATGGCCG
     1321 ACAAGCAAAA AAATGGAATC AAGGTCAACT TCAAGATTCG CCACAACATC GAAGACGGGT
     1381 CGGTTCAACT CGCTGATCAC TACCAGCAGA ACACACCAAT TGGAGgtaag ttttattcca
     1441 taatacaaat atatattgaa caataatgct tttcagATGG ACCAGTCCTC CTCCCTGATA
     1501 ATCACTACCT TTCCACTCAA TCCGCTCTTA GCAAGGATCC AAATGAGAAA AGAGATCACA
     1561 TGGTTCTTCT CGAGTTTGTC ACCGCCGCCG GAATCACCCA CGGAATGGAC GAGCTTTACA
     1621 AGtaaatgca aaatcctttc aagcattccc ttcttctcta tcactcttct ttctttttgt
     1681 caaaaaattc tctcgctaat ttatttgctt ttttaatgtt attattttat gactttttat
     1741 agtcactgaa aagtttgcat ctgagtgaag tgaatgctat caaaatgtga ttctgtctga
     1801 tgtactttca caatctctct tcaattccat tttgaagtgc tttaaacccg aaaggttgag
     1861 aaaaatgcga gcgctcaaat atttgtattg tgttcgttga gtgacccaac aaaaagagga
     1921 aactttattg tgccgccaag aaaaaagtct cacaggaaac ggagaatctg tgcggcaatg
     1981 tcttatcatt tgaaacgcgc cgtatcgatg ctgacatcat gcggctcccg tgttagtcga
     2041 gaagatgggt acgggccttt attcatccaa cactctacgg cttcttcaag agctagtagg
     2101 gcaccctgta gttggaaagg gattatttcg tatggcgagc ccataccgtc tttcttgcgg
     2161 aagcttaaca cgataggaag ttggatagtt tcgaacgatg gttccatacg gaacgctgtc
     2221 tcaaggatga gtctgacgga gtgtaactcg aaggctaggt ggaggctcag tgatgataag
     2281 tctgcgatgg tggatgcatg tgtcatggtc atagctgttt cctgtgtgaa attgttatcc
     2341 gctcagaggg cacaatccta ttccgcgcta tccgacaatc tccaagacat taggtggagt
     2401 tcagttcggc gtatggcata tgtcgctgga aagaacatgt gagcaaaagg ccagcaaaag
     2461 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2521 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2581 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2641 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2701 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2761 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2821 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2881 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     2941 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3001 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3061 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3121 ctattcaaca aagccgccgt cccgtcaagt cagcgtaaat gggtaggggg cttcaaatcg
     3181 tcctcgtgat accaattcgg agcctgcttt tttgtacaaa cttgttgata atggcaattc
     3241 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3301 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3361 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3421 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagagc cacgctcacc
     3481 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3541 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3601 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3661 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3721 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3781 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     3841 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     3901 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     3961 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4021 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4081 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4141 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4201 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4261 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cagatacctg
     4321 aaacaaaacc catcgtacgg ccaaggaagt ctccaataac tgtgatccac cacaagcgcc
     4381 agggttttcc cagtcacgac gtt   
//
