LOCUS       Exported                2210 bp ds-DNA     circular SYN 05-DEC-2018
DEFINITION  MoClo golden gate assembly DE part for gN2 gRNA (guide RNA for S. 
            pyogenes Cas9; sequence designed with NUPACK for stable tracrRNA 
            structure and open targeting region)..
ACCESSION   .
VERSION     .
KEYWORDS    UC20m
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2210)
  TITLE     CIDAR MoClo Extension
REFERENCE   2  (bases 1 to 2210)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 5, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     misc_RNA        5..24
                     /created_by="sclamons"
                     /label="gN2 target"
     misc_feature    <25..66
                     /Source="referenceToDocument: urn:local:.:h9-9u0o4st"
                     /label="Digestion of IA_UC15 extraction (concatenated
                     sequence 2)"
                     /note="Geneious type: Concatenated sequence"
     misc_RNA        25..66
                     /created_by="User"
                     /label="SP tracrRNA constant region"
     terminator      67..106
                     /created_by="User"
                     /label="tracrRNA Terminator"
     misc_feature    <67..>106
                     /Source="referenceToDocument: urn:local:.:ha-9u0o4st"
                     /label="Digestion of IA_T22 extraction (concatenated
                     sequence 3)"
                     /note="Geneious type: Concatenated sequence"
     misc_feature    107^108
                     /Original_Bases="AT"
                     /label="AT"
                     /note="Geneious type: Editing History Deletion"
     misc_feature    109..110
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     misc_feature    111..2210
                     /ApEinfo_revcolor="#6ecff6"
                     /ApEinfo_fwdcolor="#6ecff6"
                     /label="DVLA"
     primer_bind     complement(194..214)
                     /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     2106..2126
                     /Sequence="AAATAGGCGTATCACGAGGCA"
                     /Hairpin_Tm="46.5"
                     /Tm="59.6"
                     /Self_Dimer_Tm="None"
                     /%GC="47.6"
                     /modified_by="User"
                     /label="SC189/Part.Seq.FOR"
     source          1..2210
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_RNA        25..100
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    118..138
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(144..161)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(315..903)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(395..414)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1074..1934)
                     /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     1697..1716
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1935..2039)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2107..2125
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      2129..2172
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2175..2196
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 tactctcctc ctccccccca atatgtttta gagctagaaa tagcaagtta aaataaggct
       61 agtccgttat caacttgaaa aagtggcacc gagtcggtgc ttttttaggt agagacctac
      121 tagtagcggc cgctgcaggc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
      181 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
      241 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
      301 gcgttgctgg cgtttttcca caggctccgc ccccctgacg agcatcacaa aaatcgacgc
      361 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
      421 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
      481 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
      541 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
      601 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
      661 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
      721 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
      781 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
      841 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
      901 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
      961 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     1021 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     1081 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     1141 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     1201 gcaatgatac cgcgcgaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     1261 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     1321 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     1381 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     1441 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     1501 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     1561 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     1621 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     1681 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     1741 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     1801 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     1861 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     1921 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     1981 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     2041 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     2101 tataaaaata ggcgtatcac gaggcagaat ttcagataaa aaaaatcctt agctttcgct
     2161 aaggatgatt tctggaattc gcggccgctt ctagagacta gtgggtctca
//
