LOCUS       Exported                2199 bp ds-DNA     circular SYN 05-DEC-2018
DEFINITION  MoClo golden gate assembly AB part for pLbda-CasRQi (strong lambda 
            promoter with downstream dCas binding site for Qi-lab gRNA (UC16m); 
            gRNA sequence taken from DOI: 10.1016/j.cell.2013.02.022)..
ACCESSION   .
VERSION     .
KEYWORDS    P35m
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2199)
  TITLE     CIDAR MoClo Extension
REFERENCE   2  (bases 1 to 2199)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 5, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     misc_feature    1..2199
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     promoter        5..48
                     /created_by="User"
                     /label="pLambda (derived from P62)"
     misc_RNA        49..68
                     /created_by="User"
                     /label="Random spacer"
     misc_feature    69..>74
                     /Source="referenceToDocument: urn:local:.:31-9u0nbxi"
                     /label="Digestion of IA_P90 extraction (concatenated
                     sequence 1)"
                     /note="Geneious type: Concatenated sequence"
     misc_RNA        complement(69..>75)
                     /created_by="User"
                     /label="SP/ST1 Hybrid PAM"
     modified_base   75
                     /created_by="User"
                     /label="G -> C to avoid BsaI site"
     misc_RNA        complement(76..95)
                     /created_by="User"
                     /label="gRNA Target"
     misc_feature    100..2199
                     /ApEinfo_revcolor="#6ecff6"
                     /ApEinfo_fwdcolor="#6ecff6"
                     /label="DVLA"
     misc_feature    complement(101..106)
                     /Recognition_pattern="GGTCTC(1/5)"
                     /annotation_group="GGTCTC"
                     /label="BsaI"
                     /note="Geneious type: restriction site"
     primer_bind     complement(183..203)
                     /Sequence="TTACCGCCTTTGAGTGAGCTG"
                     /Hairpin_Tm="None"
                     /Tm="60.6"
                     /Self_Dimer_Tm="None"
                     /%GC="52.4"
                     /created_by="primer3"
                     /label="SC190/Part.Seq.REV"
     primer_bind     2095..2115
                     /Sequence="AAATAGGCGTATCACGAGGCA"
                     /Hairpin_Tm="46.5"
                     /Tm="59.6"
                     /Self_Dimer_Tm="None"
                     /%GC="47.6"
                     /created_by="primer3"
                     /label="SC189/Part.Seq.FOR"
     misc_feature    2180..119
                     /created_by="User"
                     /label="P35m_gib_2"
     misc_feature    2193..2198
                     /Recognition_pattern="GGTCTC(1/5)"
                     /annotation_group="GGTCTC"
                     /label="BsaI"
                     /note="Geneious type: restriction site"
     misc_feature    2193..2198
                     /Recognition_pattern="GGTCTC(1/5)"
                     /annotation_group="GGTCTC"
                     /label="BsaI"
                     /note="Geneious type: restriction site"
     source          1..2199
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    107..127
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(133..150)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(304..892)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(384..403)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1063..1923)
                     /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     1686..1705
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1924..2028)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2096..2114
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      2118..2161
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2164..2185
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 ggagctaaca ccgtgcgtgt tgactatttt acctctggcg gtgataatga tggagtccag
       61 acaccggctt tccccagacc gctaaactga aagtttacta gagacctact agtagcggcc
      121 gctgcaggct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
      181 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
      241 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
      301 gtttttccac aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      361 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      421 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      481 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      541 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      601 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      661 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      721 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
      781 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      841 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      901 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
      961 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     1021 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     1081 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     1141 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     1201 gcgcgaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     1261 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     1321 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     1381 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     1441 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     1501 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     1561 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     1621 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     1681 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     1741 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     1801 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     1861 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     1921 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     1981 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     2041 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     2101 gcgtatcacg aggcagaatt tcagataaaa aaaatcctta gctttcgcta aggatgattt
     2161 ctggaattcg cggccgcttc tagagactag tgggtctca
//
