LOCUS       sequence-121303-        3150 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Contains the sgRNA backbone sequence (tracrRNA, 82 bp) and is used 
            as DNA template to amplify a specific sgRNA using a forward primer 
            with the protospacer sequence for gene targeting..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3150)
  AUTHORS   Lander N, Li ZH, Niyogi S, Docampo R
  TITLE     CRISPR/Cas9-Induced Disruption of Paraflagellar Rod Protein 1 and 2 
            Genes in Trypanosoma cruzi Reveals Their Role in Flagellar 
            Attachment.
  JOURNAL   MBio. 2015 Jul 21;6(4). pii: e01012-15. doi: 10.1128/mBio.01012-15.
  PUBMED    26199333
REFERENCE   2  (bases 1 to 3150)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "MBio."; 
            date: "2015-07-21"; volume: "6(4). pii: e01012-15. doi"; pages: " 
            10.1128/mBio.01012-15"
COMMENT     Sequence Label: pUC_sgRNA
FEATURES             Location/Qualifiers
     source          1..3150
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(23..40)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(23..39)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(32..54)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      252..707
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(339..358)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     549..570
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(724..743)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(903..921)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        989..1093
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1094..1162,1163..1954)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1312..1331)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2125..2713
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2614..2633
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2867..2884
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3001..3022
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3037..3042,3043..3060,3061..3067)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3075..3091
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     3080..3102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3099..3115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3099..3115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gaaagttgag accatggaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
       61 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
      121 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
      181 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
      241 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
      301 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      361 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      421 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
      481 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      541 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc
      601 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      661 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
      721 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
      781 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
      841 caacgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgacccgaaa
      901 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
      961 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     1021 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     1081 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     1141 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     1201 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     1261 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     1321 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     1381 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     1441 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     1501 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     1561 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     1621 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     1681 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     1741 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     1801 tgagcgtggt tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     1861 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     1921 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     1981 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     2041 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     2101 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2161 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2221 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     2281 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2341 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2401 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     2461 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     2521 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     2581 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     2641 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     2701 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     2761 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     2821 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     2881 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     2941 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     3001 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     3061 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     3121 ttacgccaag cttggatcct atctcctttc
//
