LOCUS       pDG267_AAVS1_A3786c_attP        2368 bp    DNA     circular UNA 17-JAN-2023
DEFINITION  .
ACCESSION   urn.local...1v-hsvq857
VERSION     urn.local...1v-hsvq857
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     primer_bind     24..46
                     /Tm=56
                     /Sequence="GCAGGCTTTAAAGGAACCAATTC"
                     /Description="<html><body></body></html>"
                     /label="AVA037"
     promoter        82..330
                     /note="RNA polymerase III promoter for human U6 snRNA"
                     /label="U6 promoter"
     primer_bind     189..208
                     /%GC="35.0"
                     /Hairpin_Tm="41.5"
                     /Self_Dimer_Tm="19.2"
                     /Sequence="TACGTGACGTAGAAAGTAAT"
                     /Tm="51.5"
                     /modified_by="kzhao"
                     /label="KZ396"
     misc_feature    331..350
                     /label="spacer"
     misc_RNA        351..426
                     /note="guide RNA scaffold for the <i>Streptococcus
                     pyogenes</i> CRISPR/Cas9 system"
                     /label="gRNA scaffold"
     misc_feature    427..465
                     /label="attP"
     misc_feature    466..478
                     /label="PBS"
     misc_feature    479..515
                     /label="Editing History Insertion"
     ncRNA           479..515
                     /created_by="chenp"
                     /label="evoPQ1-trimmed"
     misc_feature    479..515
                     /Extracted_interval="473 -> 509"
                     /Original_Circular_Sequence_Length=2362
                     /label="Extracted region from Final assembled
                     Sp-pegRNA..."
     primer_bind     complement(544..567)
                     /Tm=58
                     /Sequence="GCTCACATGTCATATGTAGAGAGG"
                     /Description="<html><body></body></html>"
                     /label="AVA038"
     rep_origin      complement(619..1207)
                     /snapgene_direction="LEFT"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     misc_feature    complement(782..801)
                     /%GC="55.0"
                     /Hairpin_Tm="83.8"
                     /Self_Dimer_Tm="23.6"
                     /Sequence="ACGCTTCCCGAAGGGAGAAA"
                     /Tm="61.5"
                     /created_by="primer3"
                     /modified_by="kzhao"
                     /label="KZ397"
     primer_bind     1282..1301
                     /%GC="30.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Sequence="GAGATTATCAAAAAGGATCT"
                     /Tm="47.7"
                     /created_by="primer3"
                     /label="ACK562"
     CDS             join(complement(2170..2238),complement(1378..2169))
                     /codon_start=1
                     /gene="<i>bla</i>"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
                     /label="AmpR"
     primer_bind     1784..1803
                     /%GC="40.0"
                     /Hairpin_Tm="37.3"
                     /Self_Dimer_Tm="None"
                     /Sequence="TTGTGCAAAAAAGCGGTTAG"
                     /Tm="55.7"
                     /created_by="primer3"
                     /label="ACK563"
     promoter        complement(2239..2343)
                     /gene="<i>bla</i>"
                     /label="AmpR promoter"
     promoter        complement(2280..2308)
                     /Imported_from="<a
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla
                     sMapper</a>"
                     /Transferred_From="amp prom"
                     /Transferred_Similarity="100.00%"
                     /label="amp prom"
ORIGIN      
        1 gacgtcgcta gctgtacaaa aaagcaggct ttaaaggaac caattcagtc gactggatcc
       61 ggtaccaagg tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata
      121 cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta
      181 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta
      241 tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct
      301 ttatatatct tgtggaaagg acgaaacacc gctggccccc caccgcccca gttttagagc
      361 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
      421 cggtgcgtac accactgaga ccgcggtggt tgaccagaca aacctggcgg tggggggccg
      481 cggttctatc tagttacgcg ttaaaccaac tagaattttt ttaagcttgg gccgctcgag
      541 gtacctctct acatatgaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
      601 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
      661 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      721 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
      781 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
      841 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
      901 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
      961 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1021 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     1081 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1141 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1201 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1261 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     1321 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     1381 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     1441 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     1501 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     1561 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     1621 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     1681 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     1741 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     1801 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     1861 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     1921 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     1981 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     2041 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     2101 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     2161 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     2221 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     2281 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     2341 gcgcacattt ccccgaaaag tgccacct
//
