LOCUS       Exported                2334 bp ds-DNA     circular SYN 05-DEC-2020
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2334)
  AUTHORS   AVA
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jan 18, 2022 from SnapGene 5.0.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..2334
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     24..46
                     /label=AVA037
                     /note="color: black; sequence: GCAGGCTTTAAAGGAACCAATTC"
     promoter        82..330
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     189..208
                     /label=KZ396
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     253..272
                     /label=LKO.1 5'
                     /note="color: #a020f0; direction: RIGHT"
     misc_feature    331..350
                     /label=spacer
                     /note="color: #3366ff"
     misc_RNA        351..426
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
                     /note="color: #00ccff; direction: RIGHT"
     misc_feature    427..469
                     /label=attP
                     /note="color: #ccffcc"
     misc_feature    470..481
                     /label=PBS
                     /note="color: #339966"
     primer_bind     complement(510..533)
                     /label=AVA038
                     /note="color: black; sequence: GCTCACATGTCATATGTAGAGAGG"
     rep_origin      complement(585..1173)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     misc_feature    complement(748..767)
                     /label=KZ397
                     /note="color: #a6acb3; direction: LEFT"
     primer_bind     1248..1267
                     /label=ACK562
                     /note="color: #a020f0; direction: RIGHT"
     CDS             complement(1344..2204)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 1344 .. 2135 / #ccffcc
                        2: 2136 .. 2204 / #ccffcc / signal sequence
                     Cleavage site after base 2135"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1750..1769
                     /label=ACK563
                     /note="color: #a020f0; direction: RIGHT"
     promoter        complement(2205..2309)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     promoter        complement(2246..2274)
                     /label=amp prom
                     /note="color: #ffffff; direction: LEFT"
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 gttatggttt actccatcta gttttagagc
      361 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
      421 cggtgctacc gtacaccact gagaccgcgg tggttgacca gacaaaccta tggagtaaac
      481 ctttttttaa gcttgggccg ctcgaggtac ctctctacat atgacatgtg agcaaaaggc
      541 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
      601 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
      661 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
      721 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
      781 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
      841 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
      901 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
      961 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     1021 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     1081 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     1141 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     1201 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     1261 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     1321 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     1381 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     1441 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     1501 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     1561 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     1621 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     1681 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     1741 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     1801 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     1861 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     1921 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     1981 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     2041 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     2101 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     2161 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     2221 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     2281 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acct
//
