LOCUS       pSEM397___mosTI_        4740 bp    DNA        circular     25-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COMMENT     Unpublished reagents.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 between
            upstream of gpd-2 operon and gfp(NLS)::tbb-2 3' UTR. 
COMMENT     Use to monitor expression of transgene by co-expression of
            fluorophore. 
COMMENT     BsaI overhangs: tgcc - mcs - tgag (use  tga as the stop site in the
            BsaI overlap for transgenes)
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(2645..2661)
                     /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    3662..4627
                     /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    2833..3421
                     /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           1960..2289
                     /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    2290..2588
                     /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(1000..1126,1178..1449,1501..1690,1742..1962)
                     /locus_tag="ce-gfp(2xNLS)"
                     /label="ce-gfp(2xNLS)"
                     /ApEinfo_label="ce-gfp(2xNLS)"
                     /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"
     sig_peptide     1003..1023
                     /locus_tag="SV40"
                     /label="SV40"
                     /ApEinfo_label="SV40"
                     /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     1888..1959
                     /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"
     misc_feature    752..999
                     /locus_tag="gpd-2 operon"
                     /label="gpd-2 operon"
                     /ApEinfo_label="gpd-2 operon"
                     /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"
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 tTGAgctgtc tcatcctact ttcacctagt
      781 taactgcttg tcttaaaatc tatgcttctc tttagtatct aaaattttcc tagaagctta
      841 caagtatata aattgtctct tctcaataaa ggttgtatat ttattcatct tattgaatct
      901 gccatttcct cgtttttgcg agtttatata ccttccaatt ttctttctat tgtattttca
      961 acttctaatt ttaattcagg gaaactgctt caacgcatcA TGCCAAAGAA GAAGAGAAAG
     1021 GTGTCAAAAG GAGAAGAACT CTTTACCGGA GTCGTCCCAA TTCTCGTCGA GCTCGACGGA
     1081 GACGTAAACG GACACAAATT CTCGGTTTCC GGAGAAGGAG AAGGAGgtaa gttttacgaa
     1141 attctgaagc tcacaaaaca tggagtacat ttttcagATG CTACTTATGG AAAACTCACC
     1201 CTTAAATTCA TTTGCACCAC CGGAAAGCTC CCAGTGCCGT GGCCGACCCT TGTGACTACA
     1261 TTCTGCTACG GAGTTCAATG CTTTTCCCGC TACCCAGACC ACATGAAGCG TCACGATTTC
     1321 TTCAAATCAG CCATGCCAGA AGGATACGTC CAGGAGCGAA CAATTTTCTT CAAGGACGAT
     1381 GGAAACTACA AGACTCGTGC TGAAGTCAAG TTTGAAGGAG ATACTCTCGT GAACCGCATT
     1441 GAGCTCAAGg taagtttata atctatgatc ttaatttggc aatggaaaaa gttttttcag
     1501 GGAATCGACT TCAAAGAAGA TGGAAACATT CTTGGACACA AGCTCGAATA CAACTATAAC
     1561 TCGCACAACG TGTATATCAT GGCCGACAAG CAAAAAAATG GAATCAAGGT CAACTTCAAG
     1621 ATTCGCCACA ACATCGAAGA CGGGTCGGTT CAACTCGCTG ATCACTACCA GCAGAACACA
     1681 CCAATTGGAG gtaagtttta ttccataata caaatatata ttgaacaata atgcttttca
     1741 gATGGACCAG TCCTCCTCCC TGATAATCAC TACCTTTCCA CTCAATCCGC TCTTAGCAAG
     1801 GATCCAAATG AGAAAAGAGA TCACATGGTT CTTCTCGAGT TTGTCACCGC CGCCGGAATC
     1861 ACCCACGGAA TGGACGAGCT TTACAAGTCG AGACGCCGCA AGGCTAATCC AACCAAACTT
     1921 TCTGAAAATG CCAAGAAACT TGCCAAGGAG GTCGAGAACT AAatgcaaaa tcctttcaag
     1981 cattcccttc ttctctatca ctcttctttc tttttgtcaa aaaattctct cgctaattta
     2041 tttgcttttt taatgttatt attttatgac tttttatagt cactgaaaag tttgcatctg
     2101 agtgaagtga atgctatcaa aatgtgattc tgtctgatgt actttcacaa tctctcttca
     2161 attccatttt gaagtgcttt aaacccgaaa ggttgagaaa aatgcgagcg ctcaaatatt
     2221 tgtattgtgt tcgttgagtg acccaacaaa aagaggaaac tttattgtgc cgccaagaaa
     2281 aaagtctcac aggaaacgga gaatctgtgc ggcaatgtct tatcatttga aacgcgccgt
     2341 atcgatgctg acatcatgcg gctcccgtgt tagtcgagaa gatgggtacg ggcctttatt
     2401 catccaacac tctacggctt cttcaagagc tagtagggca ccctgtagtt ggaaagggat
     2461 tatttcgtat ggcgagccca taccgtcttt cttgcggaag cttaacacga taggaagttg
     2521 gatagtttcg aacgatggtt ccatacggaa cgctgtctca aggatgagtc tgacggagtg
     2581 taactcgaag gctaggtgga ggctcagtga tgataagtct gcgatggtgg atgcatgtgt
     2641 catggtcata gctgtttcct gtgtgaaatt gttatccgct cagagggcac aatcctattc
     2701 cgcgctatcc gacaatctcc aagacattag gtggagttca gttcggcgta tggcatatgt
     2761 cgctggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2821 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2881 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2941 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3001 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3061 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3121 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3181 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3241 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3301 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3361 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3421 agaagatcct ttgatctttt ctacggggtc tgacgctcta ttcaacaaag ccgccgtccc
     3481 gtcaagtcag cgtaaatggg tagggggctt caaatcgtcc tcgtgatacc aattcggagc
     3541 ctgctttttt gtacaaactt gttgataatg gcaattcaag gatcttcacc tagatccttt
     3601 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3661 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3721 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     3781 ccagtgctgc aatgataccg cgagagccac gctcaccggc tccagattta tcagcaataa
     3841 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     3901 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     3961 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4021 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4081 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4141 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4201 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4261 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4321 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     4381 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     4441 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4501 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     4561 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     4621 ttccgcgcac atttccccga aaagtgccag atacctgaaa caaaacccat cgtacggcca
     4681 aggaagtctc caataactgt gatccaccac aagcgccagg gttttcccag tcacgacgtt
//
