LOCUS       Exported                2220 bp ds-DNA     circular UNA 15-MAY-2018
DEFINITION  Golden Gate ligation product.
ACCESSION   urn.local...i-8rm6v0k
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2220)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, May 13, 2020 from SnapGene Viewer 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..2220
                     /organism="unspecified"
                     /mol_type="genomic DNA"
     CDS             66..1073
                     /label=Spectinomycin resistance
                     /note="/modified_by=ppuellma"
     misc_feature    complement(100..119)
                     /label=Reverse priming site [sequencing]
                     /note="/created_by=ppuellma"
     misc_feature    1406..2020
                     /label=rep - pMB1
     variation       1564
                     /label=C to T
                     /label=nonstandard type:  mutation
     misc_feature    1931..1950
                     /label=Forward priming site (sequencing
                     /note="/created_by=ppuellma"
                     /note="/modified_by=ppuellma"
     misc_feature    1931..1950
                     /label=Forward Primer_sequencing
                     /note="Geneious type: primer_bind_reverse"
                     /note="/%GC=65.0"
                     /note="/Hairpin_Tm=39.7"
                     /note="/Self_Dimer_Tm=3.2"
                     /note="/Sequence=CCTGTCGGGTTTCGCCACCT"
                     /note="/Tm=64.9"
                     /note="/created_by=primer3"
     rep_origin      2005
                     /label=ORI
     misc_feature    2039..2047
                     /label=DraIII
                     /note="Geneious type: restriction site"
                     /note="/Recognition_pattern=CACNNN^GTG"
     misc_feature    2047..2052
                     /label=BsaI
                     /note="Geneious type: restriction site"
                     /note="/Recognition_pattern=GGTCTC(1/5)"
     misc_feature    2050..2212
                     /label=TEV-His-split GFP (concatenated sequence 2)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source=referenceToDocument: urn:local:.:d-8ox8izp"
     misc_feature    2050..2057
                     /label=Geneious type: Editing History Insertion
                     /note="Geneious type: Editing History Insertion"
                     /note="/Original_Bases="
     misc_feature    2050..2057
                     /label=Bsb I cleavage site + overhang
                     /note="/created_by=ppuellma"
     misc_feature    2050..2053
                     /label=Ligation
                     /label=nonstandard type:  ligation
     misc_feature    2054..2057
                     /label=Molecular cloning overhang (TTCG)
                     /note="/created_by=ppuellma"
     misc_feature    2058..2063
                     /label=Small linker to target protein
                     /note="Geneious type: Concatenated sequence"
                     /note="/modified_by=ppuellma"
     misc_feature    2064..2084
                     /label=TEV Protease cleavage site
                     /note="/created_by=ppuellma"
     misc_feature    2082..2120
                     /label=HisTag (concatenated sequence 2)
                     /note="Geneious type: Concatenated sequence"
     misc_feature    2096
                     /label=C
                     /note="Geneious type: Editing History Replacement"
                     /note="/Original_Bases=C"
     misc_feature    2099
                     /label=C
                     /note="Geneious type: Editing History Replacement"
                     /note="/Original_Bases=C"
     misc_feature    2102
                     /label=C
                     /note="Geneious type: Editing History Replacement"
                     /note="/Original_Bases=C"
     misc_feature    2105
                     /label=C
                     /note="Geneious type: Editing History Replacement"
                     /note="/Original_Bases=C"
     misc_feature    2108
                     /label=C
                     /note="Geneious type: Editing History Replacement"
                     /note="/Original_Bases=C"
     misc_feature    2111
                     /label=C
                     /note="Geneious type: Editing History Replacement"
                     /note="/Original_Bases=C"
     misc_feature    2205..2212
                     /label=Geneious type: Editing History Insertion
                     /note="Geneious type: Editing History Insertion"
                     /note="/Original_Bases="
     misc_feature    complement(2205..2212)
                     /label=BsbI cleavage site + overhang
                     /note="/created_by=ppuellma"
     misc_feature    2205..2208
                     /label=Molecular cloning overhang (GCTT)
                     /note="/created_by=ppuellma"
     misc_feature    join(2209..2220,1..2053)
                     /label=pAGM9121 (concatenated sequence 1)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source=referenceToDocument: urn:local:.:i-7mmlal6"
     misc_feature    2209..2212
                     /label=Ligation
                     /label=nonstandard type:  ligation
     misc_feature    complement(2210..2215)
                     /label=BsaI
                     /note="Geneious type: restriction site"
                     /note="/Recognition_pattern=GGTCTC(1/5)"
ORIGIN
        1 gtgccgaggc acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt
       61 agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg
      121 cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg
      181 catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg
      241 atgttacgca gcagggcagt cgccctaaaa caaagttaaa catcatgggg gaagcggtga
      301 tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac
      361 cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca
      421 gcgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt
      481 tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg
      541 tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg
      601 aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca
      661 cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg
      721 taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc
      781 taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg
      841 tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg
      901 atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg
      961 aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt
     1021 tggaagaatt tgtccattac gtgaaaggcg agatcaccaa ggtagtcggc aaataatgtc
     1081 tagctagaaa ttcgttcaag ccgacgccgc ttcgcggcgc ggcttaactc aagcgttaga
     1141 tgcactaagc acataattgc tcacagccaa actatcaggt caagtctgct tttattattt
     1201 ttaagcgtgc ataataagcc ctacacaaat tgggagatat atcatgctgt cagaccaagt
     1261 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     1321 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     1381 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     1441 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     1501 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     1561 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     1621 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     1681 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     1741 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     1801 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     1861 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     1921 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     1981 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctgca
     2041 ctctgtggtc tcattcgggc agcgaaaatc tatattttga aggtatgggc agccatcatc
     2101 atcatcatca tagcggaatg tcggatggag ggtctggtgg cggatcaaca agccgtgacc
     2161 acatggtcct tcatgagtac gtaaatgctg ctgggattac atgagcttcg agaccacgaa
//
