LOCUS       pSEM398___mosTI_        4629 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COMMENT     Unpublished reagents.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 between
            upstream of gpd-2 operon and mScarlet::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     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(2534..2550)
                     /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    3551..4516
                     /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    2722..3310
                     /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           1849..2178
                     /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    2179..2477
                     /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..1029,1081..1285,1337..1498,1550..1851)
                     /locus_tag="ce-mScarlet"
                     /label="ce-mScarlet"
                     /ApEinfo_label="ce-mScarlet"
                     /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"
     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 TGGTTTCTAA GGGCGAAGCC
     1021 GTCATCAAGg taagttttac gaaattctga agctcacaaa acatggagta catttttcag
     1081 GAGTTCATGC GGTTCAAGGT CCACATGGAG GGTAGTATGA ACGGACACGA GTTCGAGATA
     1141 GAGGGAGAGG GAGAGGGACG GCCATACGAG GGAACCCAAA CGGCCAAGCT CAAGGTCACC
     1201 AAAGGGGGAC CACTCCCATT CTCCTGGGAT ATCCTCTCCC CACAGTTCAT GTACGGAAGT
     1261 CGTGCCTTCA CCAAGCACCC CGCAGgtaag tttataatct atgatcttaa tttggcaatg
     1321 gaaaaagttt tttcagATAT ACCAGATTAC TATAAGCAGT CCTTCCCAGA GGGATTCAAA
     1381 TGGGAGCGTG TCATGAACTT CGAGGATGGA GGAGCCGTCA CCGTCACGCA AGACACGTCC
     1441 CTCGAGGATG GAACCCTAAT CTACAAGGTA AAGCTTCGTG GAACCAACTT CCCCCCAGgt
     1501 aagttttatt ccataataca aatatatatt gaacaataat gcttttcagA TGGACCAGTC
     1561 ATGCAAAAGA AAACTATGGG ATGGGAAGCC TCGACCGAAC GTCTCTACCC AGAGGACGGG
     1621 GTCCTCAAAG GAGATATCAA GATGGCCCTA AGACTCAAGG ACGGAGGACG GTATCTTGCC
     1681 GATTTCAAGA CCACGTACAA GGCTAAGAAG CCAGTACAAA TGCCAGGGGC CTACAACGTA
     1741 GACCGTAAAC TCGACATCAC GTCCCACAAC GAGGACTACA CGGTCGTCGA ACAATACGAA
     1801 CGTTCCGAAG GACGTCACTC CACCGGAGGG ATGGACGAGC TCTACAAGTA Aatgcaaaat
     1861 cctttcaagc attcccttct tctctatcac tcttctttct ttttgtcaaa aaattctctc
     1921 gctaatttat ttgctttttt aatgttatta ttttatgact ttttatagtc actgaaaagt
     1981 ttgcatctga gtgaagtgaa tgctatcaaa atgtgattct gtctgatgta ctttcacaat
     2041 ctctcttcaa ttccattttg aagtgcttta aacccgaaag gttgagaaaa atgcgagcgc
     2101 tcaaatattt gtattgtgtt cgttgagtga cccaacaaaa agaggaaact ttattgtgcc
     2161 gccaagaaaa aagtctcaca ggaaacggag aatctgtgcg gcaatgtctt atcatttgaa
     2221 acgcgccgta tcgatgctga catcatgcgg ctcccgtgtt agtcgagaag atgggtacgg
     2281 gcctttattc atccaacact ctacggcttc ttcaagagct agtagggcac cctgtagttg
     2341 gaaagggatt atttcgtatg gcgagcccat accgtctttc ttgcggaagc ttaacacgat
     2401 aggaagttgg atagtttcga acgatggttc catacggaac gctgtctcaa ggatgagtct
     2461 gacggagtgt aactcgaagg ctaggtggag gctcagtgat gataagtctg cgatggtgga
     2521 tgcatgtgtc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc agagggcaca
     2581 atcctattcc gcgctatccg acaatctcca agacattagg tggagttcag ttcggcgtat
     2641 ggcatatgtc gctggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2701 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2761 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2821 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2881 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     2941 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3001 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3061 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3121 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3181 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3241 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3301 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctctat tcaacaaagc
     3361 cgccgtcccg tcaagtcagc gtaaatgggt agggggcttc aaatcgtcct cgtgatacca
     3421 attcggagcc tgcttttttg tacaaacttg ttgataatgg caattcaagg atcttcacct
     3481 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3541 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3601 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3661 catctggccc cagtgctgca atgataccgc gagagccacg ctcaccggct ccagatttat
     3721 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3781 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3841 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3901 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3961 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4021 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4081 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4141 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4201 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4261 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4321 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4381 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4441 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4501 aaataggggt tccgcgcaca tttccccgaa aagtgccaga tacctgaaac aaaacccatc
     4561 gtacggccaa ggaagtctcc aataactgtg atccaccaca agcgccaggg ttttcccagt
     4621 cacgacgtt     
//
