LOCUS       pSAH45____MosTI_        4387 bp    DNA        circular     22-MAR-2023
DEFINITION  .
ACCESSION   
VERSION     
SOURCE      .
  ORGANISM  .
COMMENT     MOdular Safe harbor Transgene Insertion (mosTI) (El Mouridi et al.,
            2022). Standard cloning vector for unc-119, wrmScarlet based mosTI
            insertions with MCS and Golden Gate cloning sites. Screen for
            unc-119 rescue and wrmScarlet after single-copy insertion. For more
            information visit www.wormbuilder.org
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     misc_feature    1074..1080
                     /locus_tag="MCS w 2xBsaI(transgene insert site)"
                     /label="MCS w 2xBsaI(transgene insert site)"
                     /ApEinfo_label="MCS w 2xBsaI(transgene insert site)"
                     /ApEinfo_fwdcolor="#00fa92"
                     /ApEinfo_revcolor="#00fa92"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    1262..1561
                     /locus_tag="Right repair homology"
                     /label="Right repair homology"
                     /ApEinfo_label="Right repair homology"
                     /ApEinfo_fwdcolor="#85dae9"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     CDS             4134..4240
                     /locus_tag="cbr-unc-119 (Exons 3-4)"
                     /label="cbr-unc-119 (Exons 3-4)"
                     /ApEinfo_label="cbr-unc-119 (Exons 3-4)"
                     /ApEinfo_fwdcolor="#8000ff"
                     /ApEinfo_revcolor="#8000ff"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    1562..1582
                     /locus_tag="Uni_9 R"
                     /label="Uni_9 R"
                     /ApEinfo_label="Uni_9 R"
                     /ApEinfo_fwdcolor="#cc66ff"
                     /ApEinfo_revcolor="#cc66ff"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     CDS             join(178..180,187..321,397..589,643..800,858..1064,
                     1071..1073)
                     /locus_tag="wrmScarlet(rps-20)"
                     /label="wrmScarlet(rps-20)"
                     /ApEinfo_label="wrmScarlet(rps-20)"
                     /ApEinfo_fwdcolor="#f5b5b1"
                     /ApEinfo_revcolor="#f5b5b1"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     complement(1618..1634)
                     /locus_tag="M13 rev"
                     /label="M13 rev"
                     /ApEinfo_label="M13 rev"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     3'UTR           1087..1255
                     /locus_tag="tbb-2 3'UTR"
                     /label="tbb-2 3'UTR"
                     /ApEinfo_label="tbb-2 3'UTR"
                     /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"
     rep_origin      complement(1806..2394)
                     /locus_tag="ori"
                     /label="ori"
                     /ApEinfo_label="ori"
                     /ApEinfo_fwdcolor="#b4abac"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     CDS             complement(2635..3495)
                     /locus_tag="AmpR"
                     /label="AmpR"
                     /ApEinfo_label="AmpR"
                     /ApEinfo_fwdcolor="#0080ff"
                     /ApEinfo_revcolor="#0080ff"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     3'UTR           4241..4353
                     /locus_tag="eft-3 3'UTR"
                     /label="eft-3 3'UTR"
                     /ApEinfo_label="eft-3 3'UTR"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     promoter        complement(3496..3600)
                     /locus_tag="AmpR promoter"
                     /label="AmpR promoter"
                     /ApEinfo_label="AmpR promoter"
                     /ApEinfo_fwdcolor="#ffef86"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    7..121
                     /locus_tag="Decoy sequence (start, intron, stop)"
                     /label="Decoy sequence (start, intron, stop)"
                     /ApEinfo_label="Decoy sequence (start, intron, stop)"
                     /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"
     primer_bind     3714..3730
                     /locus_tag="M13 fwd"
                     /label="M13 fwd"
                     /ApEinfo_label="M13 fwd"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    join(7..26,78..121)
                     /locus_tag="Fire Decoy (start, intron, stop)"
                     /label="Fire Decoy (start, intron, stop)"
                     /ApEinfo_label="Fire Decoy (start, intron, stop)"
                     /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    3761..3782
                     /locus_tag="Uni_9 F"
                     /label="Uni_9 F"
                     /ApEinfo_label="Uni_9 F"
                     /ApEinfo_fwdcolor="#cc66ff"
                     /ApEinfo_revcolor="#cc66ff"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    140..159
                     /locus_tag="MCS w 2xBsaI(transgene insert site)(1)"
                     /label="MCS w 2xBsaI(transgene insert site)(1)"
                     /ApEinfo_label="MCS w 2xBsaI(transgene insert site)"
                     /ApEinfo_fwdcolor="#00fa92"
                     /ApEinfo_revcolor="#00fa92"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     CDS             3783..4082
                     /locus_tag="cbr-unc-119 (Exons 3-4)(1)"
                     /label="cbr-unc-119 (Exons 3-4)(1)"
                     /ApEinfo_label="cbr-unc-119 (Exons 3-4)"
                     /ApEinfo_fwdcolor="#8000ff"
                     /ApEinfo_revcolor="#8000ff"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_recomb     4354..4387
                     /locus_tag="LoxP"
                     /label="LoxP"
                     /ApEinfo_label="LoxP"
                     /ApEinfo_fwdcolor="#d59687"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    1..6
                     /locus_tag="EcoRV - linearize"
                     /label="EcoRV - linearize"
                     /ApEinfo_label="EcoRV - linearize"
                     /ApEinfo_fwdcolor="#fd9200"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    1250..1255
                     /locus_tag="EcoRV - linearize(1)"
                     /label="EcoRV - linearize(1)"
                     /ApEinfo_label="EcoRV - linearize"
                     /ApEinfo_fwdcolor="#fa9100"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    181..186
                     /locus_tag="Apal - N-terminal fusion"
                     /label="Apal - N-terminal fusion"
                     /ApEinfo_label="Apal - N-terminal fusion"
                     /ApEinfo_fwdcolor="#fd9200"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    1065..1070
                     /locus_tag="Smal - C-terminal fusion"
                     /label="Smal - C-terminal fusion"
                     /ApEinfo_label="Smal - C-terminal fusion"
                     /ApEinfo_fwdcolor="#fa9100"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
ORIGIN
        1 gatatcaaaa ATGATTACGC CAAGCTgtaa gtttaaacat gatcttacta actaactatt
       61 ctcatttaaa ttttcagAGC TTAAAAATGG CTGAAATCAC TCACAACGAT GGATACGCTA
      121 AtgccaGAGA CGagtgcacA GCTAGCAATA CCGAGCTCAc tgcagaCGTC TCtaaaaATG
      181 GGGCCCGTTT CAAAGGGAGA GGCCGTTATA AAAGAATTTA TGCGTTTCAA GGTCCACATG
      241 GAGGGATCCA TGAACGGACA CGAGTTCGAG ATCGAGGGAG AGGGAGAGGG ACGTCCATAC
      301 GAGGGAACCC AAACCGCCAA Ggtgagtttt attatcgatt ttcgggattt tctacttgta
      361 attacctttt gtttttaata agttgctttt ttccagCTCA AGGTCACCAA GGGAGGACCA
      421 CTCCCATTCT CCTGGGACAT CCTCTCCCCA CAATTCATGT ACGGATCCCG TGCCTTCACC
      481 AAGCACCCAG CCGACATCCC AGACTACTAC AAGCAATCCT TCCCAGAGGG ATTCAAGTGG
      541 GAGCGTGTCA TGAACTTCGA GGACGGAGGA GCCGTCACCG TCACCCAAGg taggttttca
      601 gtttcaattt ttgttgcaac tacctcaact ctatgttttc agACACCTCC CTCGAGGACG
      661 GAACCCTCAT CTACAAGGTC AAGCTCCGTG GAACCAACTT CCCACCAGAC GGACCAGTCA
      721 TGCAAAAGAA GACCATGGGA TGGGAGGCCT CCACCGAGCG ACTCTACCCA GAGGACGGAG
      781 TCCTCAAGGG AGACATCAAG gtaaattttt caagcttcta aattccaaaa taaaccattt
      841 taatttctta attccagATG GCCCTCCGGC TCAAGGACGG AGGACGTTAC CTCGCCGACT
      901 TCAAGACCAC CTACAAGGCC AAGAAGCCAG TCCAAATGCC AGGAGCCTAC AACGTCGACC
      961 GTAAGCTCGA CATCACCTCC CACAACGAGG ACTACACCGT CGTCGAGCAA TACGAGCGTT
     1021 CCGAGGGACG TCACTCCACC GGAGGAATGG ACGAGCTCTA CAAGcccggg TGAtggtctc
     1081 tgagctatgc aaaatccttt caagcattcc cttcttctct atcactcttc tttctttttg
     1141 tcaaaaaatt ctctcgctaa tttatttgct tttttaatgt tattatttta tgacttttta
     1201 tagtcactga aaagtttgca tctgagtgaa gtgaatgcta tcaaaatgtg attctgatat
     1261 ctcaggaaac ggagaatctg tgcggcaatg tcttatcatt tgaaacgcgc cgtatcgatg
     1321 ctgacatcat gcggctcccg tgttagtcga gaagatgggt acgggccttt attcatccaa
     1381 cactctacgg cttcttcaag agctagtagg gcaccctgta gttggaaagg gattatttcg
     1441 tatggcgagc ccataccgtc tttcttgcgg aagcttaaca cgataggaag ttggatagtt
     1501 tcgaacgatg gttccatacg gaacgctgtc tcaaggatga gtctgacgga gtgtaactcg
     1561 aaggctaggt ggaggctcag tgatgataag tctgcgatgg tggatgcatg tgtcatggtc
     1621 atagctgttt cctgtgtgaa attgttatcc gctcagaggg cacaatccta ttccgcgcta
     1681 tccgacaatc tccaagacat taggtggagt tcagttcggc gtatggcata tgtcgctgga
     1741 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     1801 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     1861 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     1921 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     1981 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2041 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2101 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2161 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2221 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2281 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2341 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2401 cctttgatct tttctacggg gtctgacgct ctattcaaca aagccgccgt cccgtcaagt
     2461 cagcgtaaat gggtaggggg cttcaaatcg tcctcgtgat accaattcgg agcctgcttt
     2521 tttgtacaaa cttgttgata atggcaattc aaggatcttc acctagatcc ttttaaatta
     2581 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2641 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2701 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     2761 tgcaatgata ccgcgagagc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     2821 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     2881 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     2941 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3001 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3061 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3121 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3181 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3241 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3301 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3361 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3421 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     3481 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     3541 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     3601 cacatttccc cgaaaagtgc cagatacctg aaacaaaacc catcgtacgg ccaaggaagt
     3661 ctccaataac tgtgatccac cacaagcgcc agggttttcc cagtcacgac gttgtaaaac
     3721 gacggccagt catgcataat ccgcacgcat ctggaataag gaagtgccat tccgcctgac
     3781 ctAAGTGCTT TTCGAGATCG CCAAACCGGA GAACGATCAG GAGAATGATG AGTCGCCACA
     3841 GGAGTCGGCA AGATACGTGC GTTATAGATT TGCTCCAAAC TTTTTGAAAC TCAAAACGGT
     3901 CGGAGCAACT GTGGAATTCA AAGTAGGAGA CATCCCAATC CATCATTTCC GAATGATCGA
     3961 ACGTCACTTC TTCAAAGATC GCCTTCTGAA GTGTTTTGAC TTTGAATTCG GATTCTGTAT
     4021 TCCGAATTCA CGAAACAACT GTGAACATAT CTATGAGTTC CCTCAACTCT CTCAACAACT
     4081 CAgtgagtca ttattctaaa aagtacaatt caaaagacta atctcttttt cagTGGACGA
     4141 CATGATCAAC AATCCAAACG AGACTCGTTC TGACAGCTTC TATTTCGTCG ATAACAAACT
     4201 CGTCATGCAC AACAAAGCCG ACTACTCATA TGATGCATAA atcttcattg ttgagtttat
     4261 cttgttgatt tttgaataaa ttatcaactc tttacttttt aatgggttat gaaataaata
     4321 aacattgaaa actgataaac aacgttcatc tctataactt cgtatagcat acattatacg
     4381 aagttat     
//
