LOCUS       pSAH80____AWAp__        4663 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COMMENT     Alharbi & Froekjaer-Jensen, unpublished reagents.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     Green 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    join(437..503,579..743,797..948,1006..1350)
                     /locus_tag="gfp "
                     /label="gfp "
                     /ApEinfo_label="gfp "
                     /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             complement(2911..3771)
                     /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    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    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    427..432
                     /locus_tag="BsaI - exchange gfp"
                     /label="BsaI - exchange gfp"
                     /ApEinfo_label="BsaI - exchange gfp"
                     /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"
     misc_feature    1351..1357
                     /locus_tag="BsaI - exchange gfp(1)"
                     /label="BsaI - exchange gfp(1)"
                     /ApEinfo_label="BsaI - exchange gfp"
                     /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           1364..1532
                     /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    1539..1837
                     /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             4410..4516
                     /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    1838..1858
                     /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(1894..1910)
                     /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(2082..2670)
                     /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(3772..3876)
                     /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           4517..4629
                     /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     3990..4006
                     /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    4037..4058
                     /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             4059..4358
                     /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     4630..4663
                     /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    440..445
                     /locus_tag="Apal - N-terminal fusion"
                     /label="Apal - N-terminal fusion"
                     /ApEinfo_label="Apal - N-terminal fusion"
                     /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"
     misc_feature    1342..1347
                     /locus_tag="Smal - C-terminal fusion"
                     /label="Smal - C-terminal fusion"
                     /ApEinfo_label="Smal - C-terminal fusion"
                     /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"
     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    1533..1538
                     /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 GGCCCTCTAA GGGCGAAGAA TTGTTCACGG GAGTAGTCCC
      481 AATCTTAGTC GAGCTTGACG GAGgtgagtt ttattatcga ttttcgggat tttctacttg
      541 taattacctt ttgtttttaa taagttgctt ttttccagAT GTTAATGGCC ACAAGTTTTC
      601 CGTGTCTGGA GAAGGAGAAG GCGACGCTAC CTACGGTAAG CTGACCCTGA AGTTCATTTG
      661 CACTACTGGC AAGTTACCAG TACCATGGCC GACCCTGGTG ACAACATTTT GCTACGGGGT
      721 TCAATGCTTC TCCAGATACC CAGgtaggtt ttcagtttca atttttgttg caactacctc
      781 aactctatgt tttcagATCA TATGAAAAGA CACGACTTTT TTAAATCCGC CATGCCAGAG
      841 GGTTACGTTC AAGAACGTAC CATTTTTTTT AAGGACGATG GTAACTACAA GACACGTGCG
      901 GAGGTTAAAT TTGAAGGCGA TACTCTCGTG AACCGTATTG AGCTCAAGgt aaatttttca
      961 agcttctaaa ttccaaaata aaccatttta atttcttaat tccagGGAAT CGATTTCAAG
     1021 GAAGATGGAA ACATTTTGGG CCACAAGCTT GAATATAATT ATAATAGTCA TAATGTCTAC
     1081 ATTATGGCCG ACAAACAAAA AAACGGTATT AAGGTTAACT TCAAGATTCG CCACAATATC
     1141 GAAGATGGAT CTGTCCAACT TGCTGACCAC TACCAACAAA ATACTCCAAT TGGAGATGGA
     1201 CCGGTTTTGC TTCCAGATAA CCACTACCTT TCCACTCAGT CTGCCCTCTC TAAGGATCCG
     1261 AATGAAAAAC GTGACCATAT GGTTCTTTTG GAGTTCGTTA CCGCAGCAGG AATTACTCAC
     1321 GGAATGGATG AACTGTACAA GCCCGGGTGA tggtctctga gctatgcaaa atcctttcaa
     1381 gcattccctt cttctctatc actcttcttt ctttttgtca aaaaattctc tcgctaattt
     1441 atttgctttt ttaatgttat tattttatga ctttttatag tcactgaaaa gtttgcatct
     1501 gagtgaagtg aatgctatca aaatgtgatt ctgatatcca ggaaacggag aatctgtgcg
     1561 gcaatgtctt atcatttgaa acgcgccgta tcgatgctga catcatgcgg ctcccgtgtt
     1621 agtcgagaag atgggtacgg gcctttattc atccaacact ctacggcttc ttcaagagct
     1681 agtagggcac cctgtagttg gaaagggatt atttcgtatg gcgagcccat accgtctttc
     1741 ttgcggaagc ttaacacgat aggaagttgg atagtttcga acgatggttc catacggaac
     1801 gctgtctcaa ggatgagtct gacggagtgt aactcgaagg ctaggtggag gctcagtgat
     1861 gataagtctg cgatggtgga tgcatgtgtc atggtcatag ctgtttcctg tgtgaaattg
     1921 ttatccgctc agagggcaca atcctattcc gcgctatccg acaatctcca agacattagg
     1981 tggagttcag ttcggcgtat ggcatatgtc gctggaaaga acatgtgagc aaaaggccag
     2041 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2101 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2161 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2221 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2281 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2341 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2401 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2461 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2521 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2581 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2641 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2701 gacgctctat tcaacaaagc cgccgtcccg tcaagtcagc gtaaatgggt agggggcttc
     2761 aaatcgtcct cgtgatacca attcggagcc tgcttttttg tacaaacttg ttgataatgg
     2821 caattcaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2881 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2941 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3001 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagagccacg
     3061 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3121 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3181 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3241 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3301 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3361 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3421 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3481 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3541 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3601 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3661 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     3721 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3781 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3841 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccaga
     3901 tacctgaaac aaaacccatc gtacggccaa ggaagtctcc aataactgtg atccaccaca
     3961 agcgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtcatg cataatccgc
     4021 acgcatctgg aataaggaag tgccattccg cctgacctAA GTGCTTTTCG AGATCGCCAA
     4081 ACCGGAGAAC GATCAGGAGA ATGATGAGTC GCCACAGGAG TCGGCAAGAT ACGTGCGTTA
     4141 TAGATTTGCT CCAAACTTTT TGAAACTCAA AACGGTCGGA GCAACTGTGG AATTCAAAGT
     4201 AGGAGACATC CCAATCCATC ATTTCCGAAT GATCGAACGT CACTTCTTCA AAGATCGCCT
     4261 TCTGAAGTGT TTTGACTTTG AATTCGGATT CTGTATTCCG AATTCACGAA ACAACTGTGA
     4321 ACATATCTAT GAGTTCCCTC AACTCTCTCA ACAACTCAgt gagtcattat tctaaaaagt
     4381 acaattcaaa agactaatct ctttttcagT GGACGACATG ATCAACAATC CAAACGAGAC
     4441 TCGTTCTGAC AGCTTCTATT TCGTCGATAA CAAACTCGTC ATGCACAACA AAGCCGACTA
     4501 CTCATATGAT GCATAAatct tcattgttga gtttatcttg ttgatttttg aataaattat
     4561 caactcttta ctttttaatg ggttatgaaa taaataaaca ttgaaaactg ataaacaacg
     4621 ttcatctcta taacttcgta tagcatacat tatacgaagt tat 
//
