LOCUS       pTW322_Lvl1_P1          4797 bp    DNA     circular UNA 10-APR-2014
DEFINITION  No SapI or BsaI in Backbone.
ACCESSION   urn.local...jf-9e4hqdh
VERSION     urn.local...jf-9e4hqdh
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     misc_feature    118
                     /note="Geneious type: Editing History Replacement"
                     /standard_name="A"
     primer_bind     157..171
                     /standard_name="BsaXI overhang"
     terminator      complement(268..295)
                     /note="Geneious type: Terminator"
                     /standard_name="rrnB_T2 term"
     terminator      complement(427..470)
                     /note="Geneious type: Terminator"
                     /standard_name="rrnB_T1 term"
     primer_bind     469..489
                     /standard_name="TW_lvl1_seq_Fwd1"
     primer_bind     536..553
                     /standard_name="M13 (-21) Forward (Quintara)"
     primer_bind     549..596
                     /standard_name="TW_lvl1_P1_Fwd"
     primer_bind     complement(549..596)
                     /standard_name="TW_lvl1_P1_Rev"
     primer_bind     549..563
                     /standard_name="15 bp ApaI Overhang"
     misc_feature    564..570
                     /standard_name="SapI"
     misc_feature    571..581
                     /note="Geneious type: Editing History Insertion"
                     /standard_name
     misc_feature    complement(576..581)
                     /standard_name="BsaI site"
     primer_bind     582..596
                     /standard_name="15 bp ApaI Overhang"
     misc_feature    584..815
                     /note="Geneious type: attP1"
                     /standard_name="attP1"
     misc_feature    666..672
                     /standard_name="attB1 x attP1 core recombination site"
     misc_feature    816..844
                     /standard_name="Misc repeat 1 in pDONR221"
     misc_feature    843..869
                     /standard_name="Misc repeat 2 in pDONR221"
     misc_feature    864..892
                     /standard_name="Misc repeat 1 in pDONR221"
     gene            complement(1211..1516)
                     /note="Geneious type: Gene"
                     /standard_name="ccdB"
     misc_feature    complement(1296..1301)
                     /standard_name="BsaI Site"
     CDS             complement(1839..2519)
                     /standard_name="Chloramphenicol resistance gene"
     misc_feature    2740..2766
                     /standard_name="Misc repeat 2 in pDONR221"
     misc_feature    complement(2767..2998)
                     /note="Geneious type: attP2"
                     /standard_name="attP2"
     misc_feature    complement(2910..2916)
                     /standard_name="attB2 x attP2 core recombination site"
     primer_bind     2998..3049
                     /standard_name="P1_F2"
     primer_bind     complement(2998..3049)
                     /standard_name="P1_R2"
     primer_bind     2998..3012
                     /standard_name="15 bp EcoRV Overhang"
     misc_feature    3013..3034
                     /note="Geneious type: Editing History Insertion"
                     /standard_name
     misc_feature    3013..3018
                     /standard_name="BsaI"
     misc_feature    complement(3028..3034)
                     /standard_name="SapI"
     primer_bind     3035..3049
                     /standard_name="15 bp EcoRV Overhang"
     promoter        complement(3039..3057)
                     /note="Geneious type: Promoter"
                     /standard_name="T7 prom"
     primer_bind     complement(3062..3078)
                     /standard_name="M13 Reverse (Quintara)"
     primer_bind     complement(3104..3125)
                     /standard_name="TW_lvl1_seq_Rev1"
     CDS             3191..4000
                     /standard_name="Kanamycin resistance gene"
     rep_origin      4121..4794
     primer_bind     4781..4795
                     /standard_name="15 bp PciI overhang"
     primer_bind     4781..171
                     /standard_name="TW_lvl1_Sap-mut_Fwd"
     primer_bind     complement(4781..171)
                     /standard_name="TW_lvl1_Sap-mut_Rev"
ORIGIN      
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggagga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcggctcttc gactggagac cccaaataat gattttattt
      601 tgactgatag tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca
      661 actttgtaca aaaaagctga acgagaaacg taaaatgata taaatatcaa tatattaaat
      721 tagattttgc ataaaaaaca gactacataa tactgtaaaa cacaacatat ccagtcacta
      781 tgaatcaact acttagatgg tattagtgac ctgtagtcga ccgacagcct tccaaatgtt
      841 cttcgggtga tgctgccaac ttagtcgacc gacagccttc caaatgttct tctcaaacgg
      901 aatcgtcgta tccagcctac tcgctattgt cctcaatgcc gtattaaatc ataaaaagaa
      961 ataagaaaaa gaggtgcgag cctctttttt gtgtgacaaa ataaaaacat ctacctattc
     1021 atatacgcta gtgtcatagt cctgaaaatc atctgcatca agaacaattt cacaactctt
     1081 atacttttct cttacaagtc gttcggcttc atctggattt tcagcctcta tacttactaa
     1141 acgtgataaa gtttctgtaa tttctactgt atcgacctgc agactggctg tgtataaggg
     1201 agcctgacat ttatattccc cagaacatca ggttaatggc gtttttgatg tcattttcgc
     1261 ggtggctgag atcagccact tcttccccga taacggagac cggcacactg gccatatcgg
     1321 tggtcatcat gcgccagctt tcatccccga tatgcaccac cgggtaaagt tcacgggaga
     1381 ctttatctga cagcagacgt gcactggcca gggggatcac catccgtcgc ccgggcgtgt
     1441 caataatatc actctgtaca tccacaaaca gacgataacg gctctctctt ttataggtgt
     1501 aaaccttaaa ctgcatttca ccagcccctg ttctcgtcag caaaagagcc gttcatttca
     1561 ataaaccggg cgacctcagc catcccttcc tgattttccg ctttccagcg ttcggcacgc
     1621 agacgacggg cttcattctg catggttgtg cttaccagac cggagatatt gacatcatat
     1681 atgccttgag caactgatag ctgtcgctgt caactgtcac tgtaatacgc tgcttcatag
     1741 catacctctt tttgacatac ttcgggtata catatcagta tatattctta taccgcaaaa
     1801 atcagcgcgc aaatacgcat actgttatct ggcttttagt aagccggatc cacgcggcgt
     1861 ttacgccccc cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca
     1921 tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg
     1981 ccttgcgtat aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc
     2041 acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc
     2101 tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa
     2161 tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt
     2221 tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca
     2281 ccgtctttca ttgccatacg gaattccgga tgagcattca tcaggcgggc aagaatgtga
     2341 ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata
     2401 tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt
     2461 tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt
     2521 ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt
     2581 atttcattat ggtgaaagtt ggaacctctt acgtgccgat caacgtctca ttttcgccaa
     2641 aagttggccc agggcttccc ggtatcaaca gggacaccag gatttattta ttctgcgaag
     2701 tgatcttccg tcacaggtat ttattcggcg caaagtgcgt cgggtgatgc tgccaactta
     2761 gtcgactaca ggtcactaat accatctaag tagttgattc atagtgactg gatatgttgt
     2821 gttttacagt attatgtagt ctgtttttta tgcaaaatct aatttaatat attgatattt
     2881 atatcatttt acgtttctcg ttcagctttc ttgtacaaag ttggcattat aagaaagcat
     2941 tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc
     3001 catccagctg atggtctcct tcacgctgaa gagcatcccc tatagtgagt cgtattacat
     3061 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     3121 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     3181 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     3241 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     3301 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     3361 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     3421 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     3481 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     3541 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     3601 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     3661 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     3721 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     3781 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     3841 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     3901 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     3961 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     4021 gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     4081 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4141 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4201 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4261 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4321 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4381 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4441 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4501 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4561 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4621 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4681 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4741 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
