LOCUS       pSAH79____AWAp__        4645 bp    DNA        circular     02-JUL-2024
DEFINITION  .
ACCESSION   
VERSION     
SOURCE      .
  ORGANISM  .
COMMENT     Alharbi & Froekjaer-Jensen, unpublished reagents. For details, see
            www.wormbuilder.org/BioPart
COMMENT     Red fluorescence marker and cloning vector for expression in the
            AWA neurons. 
COMMENT     The vector is compatible with single-copy insertions using MosTI
            (El Mouridi et al., 2022) but can also be used in extra-chromosomal
            arrays. 
COMMENT     Verified expression in AWA in hermaphrodites and males in
            extrachromosomal arrays and visible expression on fluorescence
            dissection microscope from single-copy insertions. 
COMMENT     Use BsaI to exchange the fluorophore for a different transgene
            (AAAA and GAGC Golden-gate overhangs).
COMMENT     Use ApaI for N-terminal fusions
COMMENT     Use SmaI for C-terminal fusions
COMMENT     Use EcoRV to linearize the construct and remove MosTI sequences.
COMMENT     Sequence annotations are optimized for use with ApE: A Plasmid
            Editor (https://doi.org/10.3389/fbinf.2022.818619)
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     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"
     misc_feature    126..421
                     /locus_tag="AWBp (WBGene00006109, str-44)"
                     /label="AWBp (WBGene00006109, str-44)"
                     /ApEinfo_label="AWBp (WBGene00006109, str-44)"
                     /ApEinfo_fwdcolor="#f3d46c"
                     /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    427..432
                     /locus_tag="BsaI - exchange mScarlet"
                     /label="BsaI - exchange mScarlet"
                     /ApEinfo_label="BsaI - exchange mScarlet"
                     /ApEinfo_fwdcolor="#fe9200"
                     /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    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"
     CDS             join(437..439,446..580,656..848,902..1059,1117..1323,
                     1330..1332)
                     /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"
     CDS             complement(2893..3753)
                     /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"
     misc_feature    1333..1339
                     /locus_tag="BsaI - exchange wrmScarlet"
                     /label="BsaI - exchange wrmScarlet"
                     /ApEinfo_label="BsaI - exchange wrmScarlet"
                     /ApEinfo_fwdcolor="#fe9200"
                     /ApEinfo_revcolor="#00fa92"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     3'UTR           1346..1514
                     /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"
     misc_feature    1521..1819
                     /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             4392..4498
                     /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    1820..1840
                     /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"
     primer_bind     complement(1876..1892)
                     /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"
     rep_origin      complement(2064..2652)
                     /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"
     promoter        complement(3754..3858)
                     /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"
     3'UTR           4499..4611
                     /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"
     primer_bind     3972..3988
                     /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    4019..4040
                     /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"
     CDS             4041..4340
                     /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     4612..4645
                     /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="#fa9225"
                     /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    440..445
                     /locus_tag="Apal - N-terminal fusion"
                     /label="Apal - N-terminal fusion"
                     /ApEinfo_label="Apal - N-terminal fusion"
                     /ApEinfo_fwdcolor="#fe9200"
                     /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    1324..1329
                     /locus_tag="Smal - C-terminal fusion"
                     /label="Smal - C-terminal fusion"
                     /ApEinfo_label="Smal - C-terminal fusion"
                     /ApEinfo_fwdcolor="#fe9200"
                     /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    1515..1520
                     /locus_tag="EcoRV - linearize(1)"
                     /label="EcoRV - linearize(1)"
                     /ApEinfo_label="EcoRV - linearize"
                     /ApEinfo_fwdcolor="#fd9101"
                     /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 Atgccaaatg taacatcttt cattgttcat aaaaatatac gtgttcgata gttgagtttc
      181 atgagtcaac aattagaact aagtcagaat gttttgaaat ttttgaaata aaaagctacg
      241 attcaataac tcctatcttg attttatgat tttgacagaa ccgtcatcat tctcattact
      301 cattttattc ttttcgtgtg gaaatttcat gaaaaacaac tacagaaaaa actacaaatt
      361 ttgttcttaa agtaacacaa taaaagggaa ttcttagtgg gattttagtc acacttccga
      421 gaaaaagaga ccaaaaATGG GGCCCGTTTC AAAGGGAGAG GCCGTTATAA AAGAATTTAT
      481 GCGTTTCAAG GTCCACATGG AGGGATCCAT GAACGGACAC GAGTTCGAGA TCGAGGGAGA
      541 GGGAGAGGGA CGTCCATACG AGGGAACCCA AACCGCCAAG gtgagtttta ttatcgattt
      601 tcgggatttt ctacttgtaa ttaccttttg tttttaataa gttgcttttt tccagCTCAA
      661 GGTCACCAAG GGAGGACCAC TCCCATTCTC CTGGGACATC CTCTCCCCAC AATTCATGTA
      721 CGGATCCCGT GCCTTCACCA AGCACCCAGC CGACATCCCA GACTACTACA AGCAATCCTT
      781 CCCAGAGGGA TTCAAGTGGG AGCGTGTCAT GAACTTCGAG GACGGAGGAG CCGTCACCGT
      841 CACCCAAGgt aggttttcag tttcaatttt tgttgcaact acctcaactc tatgttttca
      901 gACACCTCCC TCGAGGACGG AACCCTCATC TACAAGGTCA AGCTCCGTGG AACCAACTTC
      961 CCACCAGACG GACCAGTCAT GCAAAAGAAG ACCATGGGAT GGGAGGCCTC CACCGAGCGA
     1021 CTCTACCCAG AGGACGGAGT CCTCAAGGGA GACATCAAGg taaatttttc aagcttctaa
     1081 attccaaaat aaaccatttt aatttcttaa ttccagATGG CCCTCCGGCT CAAGGACGGA
     1141 GGACGTTACC TCGCCGACTT CAAGACCACC TACAAGGCCA AGAAGCCAGT CCAAATGCCA
     1201 GGAGCCTACA ACGTCGACCG TAAGCTCGAC ATCACCTCCC ACAACGAGGA CTACACCGTC
     1261 GTCGAGCAAT ACGAGCGTTC CGAGGGACGT CACTCCACCG GAGGAATGGA CGAGCTCTAC
     1321 AAGCCCGGGT GAtggtctct gagctatgca aaatcctttc aagcattccc ttcttctcta
     1381 tcactcttct ttctttttgt caaaaaattc tctcgctaat ttatttgctt ttttaatgtt
     1441 attattttat gactttttat agtcactgaa aagtttgcat ctgagtgaag tgaatgctat
     1501 caaaatgtga ttctgatatc caggaaacgg agaatctgtg cggcaatgtc ttatcatttg
     1561 aaacgcgccg tatcgatgct gacatcatgc ggctcccgtg ttagtcgaga agatgggtac
     1621 gggcctttat tcatccaaca ctctacggct tcttcaagag ctagtagggc accctgtagt
     1681 tggaaaggga ttatttcgta tggcgagccc ataccgtctt tcttgcggaa gcttaacacg
     1741 ataggaagtt ggatagtttc gaacgatggt tccatacgga acgctgtctc aaggatgagt
     1801 ctgacggagt gtaactcgaa ggctaggtgg aggctcagtg atgataagtc tgcgatggtg
     1861 gatgcatgtg tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcagagggca
     1921 caatcctatt ccgcgctatc cgacaatctc caagacatta ggtggagttc agttcggcgt
     1981 atggcatatg tcgctggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     2041 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     2101 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2161 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2221 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2281 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2341 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2401 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2461 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     2521 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2581 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     2641 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctct attcaacaaa
     2701 gccgccgtcc cgtcaagtca gcgtaaatgg gtagggggct tcaaatcgtc ctcgtgatac
     2761 caattcggag cctgcttttt tgtacaaact tgttgataat ggcaattcaa ggatcttcac
     2821 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2881 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2941 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3001 accatctggc cccagtgctg caatgatacc gcgagagcca cgctcaccgg ctccagattt
     3061 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3121 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3181 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     3241 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     3301 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     3361 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     3421 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     3481 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     3541 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     3601 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     3661 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     3721 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     3781 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     3841 acaaataggg gttccgcgca catttccccg aaaagtgcca gatacctgaa acaaaaccca
     3901 tcgtacggcc aaggaagtct ccaataactg tgatccacca caagcgccag ggttttccca
     3961 gtcacgacgt tgtaaaacga cggccagtca tgcataatcc gcacgcatct ggaataagga
     4021 agtgccattc cgcctgacct AAGTGCTTTT CGAGATCGCC AAACCGGAGA ACGATCAGGA
     4081 GAATGATGAG TCGCCACAGG AGTCGGCAAG ATACGTGCGT TATAGATTTG CTCCAAACTT
     4141 TTTGAAACTC AAAACGGTCG GAGCAACTGT GGAATTCAAA GTAGGAGACA TCCCAATCCA
     4201 TCATTTCCGA ATGATCGAAC GTCACTTCTT CAAAGATCGC CTTCTGAAGT GTTTTGACTT
     4261 TGAATTCGGA TTCTGTATTC CGAATTCACG AAACAACTGT GAACATATCT ATGAGTTCCC
     4321 TCAACTCTCT CAACAACTCA gtgagtcatt attctaaaaa gtacaattca aaagactaat
     4381 ctctttttca gTGGACGACA TGATCAACAA TCCAAACGAG ACTCGTTCTG ACAGCTTCTA
     4441 TTTCGTCGAT AACAAACTCG TCATGCACAA CAAAGCCGAC TACTCATATG ATGCATAAat
     4501 cttcattgtt gagtttatct tgttgatttt tgaataaatt atcaactctt tactttttaa
     4561 tgggttatga aataaataaa cattgaaaac tgataaacaa cgttcatctc tataacttcg
     4621 tatagcatac attatacgaa gttat   
//
