LOCUS       pSAH72____AWAp__        4664 bp    DNA        circular     22-MAR-2023
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COMMENT     Alharbi & Froekjaer-Jensen, unpublished reagents.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(2912..3772)
                     /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..1838
                     /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             4411..4517
                     /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    1839..1859
                     /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(1895..1911)
                     /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(2083..2671)
                     /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(3773..3877)
                     /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           4518..4630
                     /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     3991..4007
                     /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    4038..4059
                     /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             4060..4359
                     /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     4631..4664
                     /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="AWAp (WBGene00003856, odr-10)"
                     /label="AWAp (WBGene00003856, odr-10)"
                     /ApEinfo_label="AWAp (WBGene00003856, odr-10)"
                     /ApEinfo_fwdcolor="#f7d470"
                     /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 Atgccattgg aataataaat tcaatatatt ttcccaattc tgaatttgtg gatacaaacg
      181 tattattttt aactgaatac ctcgtgaaat cagatttcag attttgaaaa caatatatct
      241 gactataagt tgcgtatgta gacaaattcc aggaaaataa tggctcaaac gacacgcccc
      301 tggcttcaag ttactataaa ataggttagg aacaaagcac atataaccat gtattcagtt
      361 taattaattt tttgttggcg gtggcatgaa attctcataa tttcattaag ataccttaca
      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 ctgatatctc aggaaacgga gaatctgtgc
     1561 ggcaatgtct tatcatttga aacgcgccgt atcgatgctg acatcatgcg gctcccgtgt
     1621 tagtcgagaa gatgggtacg ggcctttatt catccaacac tctacggctt cttcaagagc
     1681 tagtagggca ccctgtagtt ggaaagggat tatttcgtat ggcgagccca taccgtcttt
     1741 cttgcggaag cttaacacga taggaagttg gatagtttcg aacgatggtt ccatacggaa
     1801 cgctgtctca aggatgagtc tgacggagtg taactcgaag gctaggtgga ggctcagtga
     1861 tgataagtct gcgatggtgg atgcatgtgt catggtcata gctgtttcct gtgtgaaatt
     1921 gttatccgct cagagggcac aatcctattc cgcgctatcc gacaatctcc aagacattag
     1981 gtggagttca gttcggcgta tggcatatgt cgctggaaag aacatgtgag caaaaggcca
     2041 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2101 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2161 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2221 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2281 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2341 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2401 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2461 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2521 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2581 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2641 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2701 tgacgctcta ttcaacaaag ccgccgtccc gtcaagtcag cgtaaatggg tagggggctt
     2761 caaatcgtcc tcgtgatacc aattcggagc ctgctttttt gtacaaactt gttgataatg
     2821 gcaattcaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     2881 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     2941 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3001 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagagccac
     3061 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3121 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3181 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3241 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3301 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3361 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     3421 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3481 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     3541 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     3601 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     3661 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     3721 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     3781 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     3841 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccag
     3901 atacctgaaa caaaacccat cgtacggcca aggaagtctc caataactgt gatccaccac
     3961 aagcgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtcat gcataatccg
     4021 cacgcatctg gaataaggaa gtgccattcc gcctgacctA AGTGCTTTTC GAGATCGCCA
     4081 AACCGGAGAA CGATCAGGAG AATGATGAGT CGCCACAGGA GTCGGCAAGA TACGTGCGTT
     4141 ATAGATTTGC TCCAAACTTT TTGAAACTCA AAACGGTCGG AGCAACTGTG GAATTCAAAG
     4201 TAGGAGACAT CCCAATCCAT CATTTCCGAA TGATCGAACG TCACTTCTTC AAAGATCGCC
     4261 TTCTGAAGTG TTTTGACTTT GAATTCGGAT TCTGTATTCC GAATTCACGA AACAACTGTG
     4321 AACATATCTA TGAGTTCCCT CAACTCTCTC AACAACTCAg tgagtcatta ttctaaaaag
     4381 tacaattcaa aagactaatc tctttttcag TGGACGACAT GATCAACAAT CCAAACGAGA
     4441 CTCGTTCTGA CAGCTTCTAT TTCGTCGATA ACAAACTCGT CATGCACAAC AAAGCCGACT
     4501 ACTCATATGA TGCATAAatc ttcattgttg agtttatctt gttgattttt gaataaatta
     4561 tcaactcttt actttttaat gggttatgaa ataaataaac attgaaaact gataaacaac
     4621 gttcatctct ataacttcgt atagcataca ttatacgaag ttat 
//
