LOCUS       pAGK43                  6932 bp    DNA     circular UNA 29-MAY-2024
DEFINITION  112bp_donor-Eco1, PAM 19.
ACCESSION   urn.local...1ra-e5vigdg
VERSION     urn.local...1ra-e5vigdg
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     promoter        1..364
                     /gene="CAG Enhancer"
                     /standard_name="CAG enhancer"
     promoter        365..646
                     /gene="CAG Promoter"
                     /standard_name="CAG promoter"
     promoter        647..1525
                     /gene="CAG Intron"
                     /standard_name="CAG intron"
     motif           complement(1221..1227)
                     /created_by="katecrawford"
                     /label="PaqCI site"
     misc_feature    1615..1732
                     /created_by="katecrawford"
                     /label="MCS"
     misc_feature    1733..>1738
                     /standard_name="Eco3_GG_insert (concatenated sequence 2)"
                     /note="Geneious type: Concatenated sequence"
     misc_feature    <1733..1736
                     /standard_name="pSCL273 (concatenated sequence 1)"
                     /note="Geneious type: Concatenated sequence"
     misc_feature    1733..1736
                     /created_by="katecrawford"
                     /label="lig 1"
     motif           1733..1738
                     /created_by="katecrawford"
                     /label="Kozak"
     gene            1739..>2455
                     /created_by="Seth"
                     /modified_by="Seth"
                     /label="EGFP"
     misc_feature    <1739..>2455
                     /standard_name="Eco3_GG_insert (concatenated sequence 2)"
                     /note="Geneious type: Concatenated sequence"
     misc_feature    <2456..>4118
                     /standard_name="Eco3_GG_insert (concatenated sequence 2)"
                     /note="Geneious type: Concatenated sequence"
     motif           2465..2521
                     /created_by="User"
                     /label="P2A"
     CDS             <2522..>3481
                     /note="Derived using Geneious Prime 2021.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="ec86RT (mOpt)"
     misc_feature    <2522..>3481
                     /standard_name="Eco3_GG_insert (concatenated sequence 2)"
                     /note="Geneious type: Concatenated sequence"
     misc_feature    <3482..>3505
                     /standard_name="Eco3_GG_insert (concatenated sequence 2)"
                     /note="Geneious type: Concatenated sequence"
     motif           3482..3505
                     /created_by="katecrawford"
                     /label="SV40 NLS"
     polyA_signal    3533..3757
                     /note="bovine growth hormone polyadenylation signal"
                     /standard_name="bGH poly(A) signal"
     promoter        3792..4006
                     /note="human H1 RNA promoter"
                     /standard_name="H1 promoter"
     gene            <4011..>4118
                     /standard_name="ncRNA"
     motif           4011..4025
                     /created_by="katecrawford"
                     /label="extended a1"
     motif           4026..4037
                     /standard_name="endogenous a1"
     motif           4038
                     /standard_name="priming G"
     misc_RNA        4041..4050
                     /standard_name="P2"
     misc_RNA        4051..4056
                     /standard_name="P2 loop"
     misc_RNA        4057..4067
                     /standard_name="P2"
     misc_RNA        4068..4075
                     /standard_name="P3"
     misc_RNA        4076..4078
                     /standard_name="P3 loop"
     misc_RNA        4079..4086
                     /standard_name="P3"
     misc_feature    complement(4097..>4118)
                     /standard_name="RT-DNA"
     misc_RNA        4111..4118
                     /standard_name="possible stem base"
     misc_RNA        4113..4118
                     /standard_name="canonical stem base"
     donor           4123..4234
                     /created_by="asim.khan"
                     /modified_by="asim.khan"
                     /label="112 bp donor"
     gRNA            4165..4182
                     /created_by="asim.khan"
     edit            4179
                     /created_by="asim.khan"
                     /modified_by="asim.khan"
                     /label="Middle Edit (T>A)"
     misc_RNA        4183..>4185
                     /created_by="Seth"
                     /modified_by="Seth"
                     /modified_by="asim.khan"
                     /label="19PAM recode GG> AT"
     ncRNA           4235..4256
                     /created_by="asim.khan"
                     /label="right ncRNA"
     motif           complement(4257..4283)
                     /created_by="katecrawford"
                     /label="Extended a1/a2"
     misc_RNA        4284..4301
                     /created_by="katecrawford"
                     /label="gRNA"
     misc_feature    4302..4392
                     /standard_name="modified SpCas9 scaffold"
     misc_feature    complement(4393..4399)
                     /standard_name="term"
     misc_feature    complement(4400..4424)
                     /standard_name="hom"
     promoter        complement(4422..4543)
                     /note="Derived using Geneious Prime 2020.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="Derived using Geneious Prime 2021.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="pLac"
     Promoter        complement(4466..4495)
                     /standard_name="lac prom"
     oligo           4620..4639
                     /note="Derived using Geneious Prime 2021.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="SV40Promoter(20mer)"
     rep_origin      4636..4771
     terminator      4791..4912
                     /note="simian vacuolating virus 40 (SV40) polyA signal"
                     /regulatory_class="terminator"
                     /note="Derived using Geneious Prime 2020.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="Derived using Geneious Prime 2021.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="SV40 polyA"
     gene            complement(5923..6783)
                     /note="Derived using Geneious Prime 2021.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="AmpR"
     promoter        complement(6784..6888)
                     /note="Derived using Geneious Prime 2021.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="AmpR promoter"
     Promoter        complement(6825..6853)
                     /standard_name="amp prom"
ORIGIN      
        1 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
       61 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      121 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      181 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      241 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      301 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      361 accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca
      421 cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg
      481 gggggggggg gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg
      541 aaaggtgcgg cggcagccaa tcaaagcggc gcgctccgaa agtttccttt tatggcgagg
      601 cggcggcggc ggcggcccta taaaaagcga accgcgcggc gggcgggagt cgctgcgcgc
      661 tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg
      721 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattac
      781 cgcttggttt aatgacggct tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc
      841 cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg
      901 gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg
      961 gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg
     1021 tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga
     1081 gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc tccccgagtt
     1141 gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc
     1201 gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc
     1261 ggggagggct cgggggaggg gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg
     1321 gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg
     1381 tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc
     1441 ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg
     1501 ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg ggacggctgc
     1561 cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga
     1621 gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg
     1681 gttattgtgc tgtctcatca ttttggcaaa gaattgagct cgatatcggt acgccaccat
     1741 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1801 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     1861 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1921 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     1981 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     2041 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     2101 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     2161 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     2221 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     2281 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     2341 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     2401 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gatgagctgt acaagggaag
     2461 cggagctact aacttcagcc tgctgaagca ggctggagac gtggaggaga accctggacc
     2521 tatgaaatct gcagagtatc tgaatacgtt ccgccttagg aatttgggcc tccccgtgat
     2581 gaacaatctc cacgatatga gcaaggcgac tcgaatatcc gtggaaacgc tgagactgct
     2641 catctataca gcagactttc ggtacaggat ctacacggtc gaaaagaagg ggcctgagaa
     2701 acgcatgcga acaatttatc aacctagccg agagctcaag gcgttgcagg gctgggttct
     2761 tcgaaacatc cttgacaaac tctcatcatc accctttagt attgggtttg aaaagcacca
     2821 aagcatcctt aacaacgcga cgccacacat aggtgccaat ttcatattga acatcgactt
     2881 ggaggatttt tttccgagcc tcacagccaa taaagtgttc ggtgtttttc acagtcttgg
     2941 gtacaatcgc cttattagtt ccgttcttac caagatttgt tgttacaaga atctcttgcc
     3001 ccagggagca cccagcagtc cgaaattggc gaatttgatt tgttccaagc tcgattatcg
     3061 aatacaaggg tacgcgggca gccggggact catctatacc cgctacgcag acgatcttac
     3121 gctgtctgcc caatcaatga agaaggtcgt aaaggcgcgg gatttcttgt tttctatcat
     3181 cccgtccgag ggcttggtaa ttaattccaa aaagacttgt atctcaggac cacgatctca
     3241 gcgaaaagtg acaggactcg tcatttctca agaaaaagtc ggtataggga gagagaagta
     3301 taaggaaatc cgcgcgaaga tccaccacat attctgtggc aagagcagcg agatagaaca
     3361 cgtccgaggc tggttgtcct tcatactgag cgtggactca aaaagccacc gccggttgat
     3421 cacctatatt tcaaaactgg aaaagaaata tggaaagaac ccactcaaca aagctaaaac
     3481 accaccaaag aagaaaagaa aggtctgatc ccggttagta atgagtttaa acctgtgcct
     3541 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3601 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3661 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3721 aatagcaggc atgctgggga tgcggtgggc tctatggaga tctgcgaaat cggatccagc
     3781 tgaatctaag tgaacgctga cgtcatcaac ccgctccaag gaatcgcggg cccagtgtca
     3841 ctaggcggga acacccagcg cgcgtgcgcc ctggcaggaa gatggctgtg agggacaggg
     3901 gagtggcgcc ctgcaatatt tgcatgtcgc tatgtgttct gggaaatcac cataaacgtg
     3961 aaatgtcttt ggatttggga atcttataag ttctgtatga gaccacgaaa tgataagatt
     4021 ccgtatgcgc acccttagcg agaggtttat cattaaggtc aacctctgga tgttgtttcg
     4081 gcatcctgca ttgaatctga gttactgtct gttttccttc ctacacacct cgtgactgtc
     4141 tacgccacgt aggacacatg gttatctggg gtaaaggcag tacatcatcc ctccattgtt
     4201 cactctgggg gttcatcacc tcaaccatta tgtcaggaaa cccgtttctt ctgacgtaag
     4261 ggtgcgcata cggaatctta tcatctgggg taaaggctgt agtttcagag ctatgctgga
     4321 aacagcatag caagttgaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag
     4381 tcggtgcttt ttttttttta gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4441 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4501 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4561 gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt agtcagcaac catagtcccg
     4621 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4681 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     4741 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct aacttgttta
     4801 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     4861 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     4921 ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4981 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5041 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaagaaa
     5101 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5161 gtttttccat atgctccgcc cccctgacga gcatcacaaa aatcgacgct caggtcatag
     5221 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5281 gcgctctcct gttccaaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5341 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5401 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5461 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5521 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5581 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5641 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5701 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5761 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5821 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5881 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5941 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6001 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6061 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6121 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6181 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6241 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6301 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6361 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6421 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6481 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6541 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6601 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6661 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6721 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6781 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6841 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6901 aaaagtgcca cctgggtcga cattgattat tg
//
