LOCUS       Exported                3193 bp ds-DNA     circular SYN 11-MAY-2018
DEFINITION  SapTrap destination vector for building sgRNA expression vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3193)
  AUTHORS   Schwartz ML, Jorgensen EM
  TITLE     SapTrap, a Toolkit for High-Throughput CRISPR/Cas9 Gene Modification
            in Caenorhabditis elegans.
  JOURNAL   Genetics. 2016 Feb 2. pii: genetics.115.184275.
  PUBMED    26837755
REFERENCE   2  (bases 1 to 3193)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jun 1, 2020 from SnapGene Viewer 5.1.3
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genetics. 
            2016 Feb 2. pii: genetics.115.184275."
FEATURES             Location/Qualifiers
     source          1..3193
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1125..3193
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label=source:synthetic DNA construct
                     /note="/locus_tag=source:synthetic DNA construct"
     primer_bind     complement(29..51)
                     /locus_tag="pGEX 3'"
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /locus_tag="pRS-marker"
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectablemarker"
     primer_bind     364..386
                     /locus_tag="M13/pUC Forward"
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /locus_tag="M13 Forward"
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 
                     (-21)Forward"
     primer_bind     379..395
                     /locus_tag="M13 fwd"
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        402..749
                     /locus_tag="U6 Promoter(1)"
                     /label=U6 Promoter
                     /label=U6 Promoter(1)
     enhancer        complement(462..469)
                     /locus_tag="Putative Enhancer"
                     /label=Putative Enhancer
     misc_feature    750..752
                     /locus_tag="TTG overlap"
                     /label=TTG overlap
     promoter        750..751
                     /locus_tag="U6 Promoter"
                     /label=U6 Promoter
     misc_RNA        752
                     /locus_tag="gRNA"
                     /label=gRNA
     misc_feature    753..760
                     /locus_tag="SapI Insertion Site(1)"
                     /label=SapI Insertion Site
                     /label=SapI Insertion Site(1)
     misc_feature    771..778
                     /locus_tag="SapI Insertion Site"
                     /label=SapI Insertion Site
     misc_feature    779..871
                     /locus_tag="sgRNA"
                     /label=sgRNA
     terminator      872..881
                     /locus_tag="U6 Terminator"
                     /label=U6 Terminator
     misc_feature    882..947
                     /locus_tag="lacZ_a"
                     /label=lacZ_a
                     /note="/translation=MGKIQKVGRWVPHELNERQMERRKNTCEILLSRYKRKSF
                     LHRIV 
                     TGDEKWIFFVNPKRKKSYVDPGQPATSTARPNRFGKKTMLCVWWDQSGVIYYELLKPG 
                     ETVNTARYQQQLINLNRALQRKRPEYQKRQHRVIFLHDNAPSHTARAVRDTLETLNWE 
                     VLPHAAYSPDLAPSDYHLFASMGHALAEQRFDSYESVKKWLDEWFAAKDDEFYWRGIH 
                     KLPERWEKCVASDGKYFE*"
     primer_bind     882..901
                     /locus_tag="U6 Vector 3' #162"
                     /label=U6 Vector 3' #162
     CDS             883..951
                     /locus_tag="LacZ alpha"
                     /label=LacZ alpha
     primer_bind     complement(970..986)
                     /locus_tag="M13 rev"
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(970..986)
                     /locus_tag="M13 Reverse"
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(983..1005)
                     /locus_tag="M13/pUC Reverse"
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    994..1010
                     /locus_tag="lac operator"
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator 
                     toinhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(1018..1048)
                     /locus_tag="lac promoter"
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1063..1084
                     /locus_tag="CAP binding site"
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the 
                     presenceof cAMP."
     primer_bind     complement(1203..1220)
                     /locus_tag="L4440"
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1374..1962)
                     /direction=LEFT
                     /locus_tag="ori"
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1454..1473)
                     /locus_tag="pBR322ori-F"
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2133..2993)
                     /codon_start=1
                     /gene="bla"
                     /locus_tag="AmpR"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin,and 
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2756..2775
                     /locus_tag="Amp-R"
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2994..3098)
                     /gene="bla"
                     /locus_tag="AmpR promoter"
                     /label=AmpR promoter
     primer_bind     3166..3184
                     /locus_tag="pBRforEco"
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, 
                     forwardprimer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt ccaaaaaaaa ctagcaataa
      421 aggaataaaa aactgtacac cttaaaggcg cacactctgt tttgcaaatt ttatttttag
      481 ttgtgaattt tctgctgaga cctgaaaata gcaactttag tactactata atttgtcaac
      541 cttttcaaaa aaagcatgca atttttgaga aactcttata aaagctatta ttaaaaaaac
      601 accttttttc caaaattatt ccacaaaaaa tatgttatga aatgcctaca ccctctcaca
      661 cacactcttt atactactct gtcaaactca cgagatgtct gccgcctctt gtgttgcccc
      721 tatataaaca cctcctattg cgagatgtct tgcgaagagc ggatccatgg gctcttcggt
      781 ttaagagcta tgctggaaac agcatagcaa gtttaaataa ggctagtccg ttatcaactt
      841 gaaaaagtgg caccgagtcg gtgctttttt ttgtgaaatt tctggcgtaa tagcgaagag
      901 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gaagcttggc
      961 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     1021 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     1081 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     1141 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct catatgcgct
     1201 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     1261 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     1321 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1381 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     1441 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     1501 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     1561 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     1621 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     1681 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     1741 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     1801 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     1861 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     1921 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     1981 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2041 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     2101 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     2161 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     2221 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     2281 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     2341 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     2401 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     2461 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     2521 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     2581 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     2641 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     2701 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     2761 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     2821 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     2881 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     2941 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     3001 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     3061 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     3121 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     3181 aggccctttc gtc
//
