LOCUS       pTW328_Lvl1_P3          4813 bp    DNA     circular UNA 10-APR-2014
DEFINITION  No SapI or BsaI in Backbone.
ACCESSION   urn.local...jl-9e4hqdi
VERSION     urn.local...jl-9e4hqdi
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     misc_feature    118
                     /note="Geneious type: Editing History Replacement"
                     /standard_name="A"
     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     536..553
                     /standard_name="M13 (-21) Forward (Quintara)"
     primer_bind     549..615
                     /standard_name="P3_F1"
     primer_bind     complement(549..615)
                     /standard_name="P3_R1"
     primer_bind     549..563
                     /standard_name="15 bp ApaI Overhang"
     misc_feature    564..570
                     /standard_name="SapI"
     misc_feature    571..600
                     /note="Geneious type: Editing History Insertion"
                     /standard_name
     primer_bind     572..589
                     /standard_name="Quint BGH"
     misc_feature    complement(595..600)
                     /standard_name="BsaI site"
     primer_bind     601..615
                     /standard_name="15 bp ApaI Overhang"
     misc_feature    603..834
                     /note="Geneious type: attP1"
                     /standard_name="attP1"
     misc_feature    685..691
                     /standard_name="attB1 x attP1 core recombination site"
     misc_feature    835..863
                     /standard_name="Misc repeat 1 in pDONR221"
     misc_feature    862..888
                     /standard_name="Misc repeat 2 in pDONR221"
     misc_feature    883..911
                     /standard_name="Misc repeat 1 in pDONR221"
     gene            complement(1230..1535)
                     /note="Geneious type: Gene"
                     /standard_name="ccdB"
     misc_feature    complement(1315..1320)
                     /standard_name="BsaI Site"
     CDS             complement(1858..2538)
                     /standard_name="Chloramphenicol resistance gene"
     misc_feature    2759..2785
                     /standard_name="Misc repeat 2 in pDONR221"
     misc_feature    complement(2786..3017)
                     /note="Geneious type: attP2"
                     /standard_name="attP2"
     misc_feature    complement(2929..2935)
                     /standard_name="attB2 x attP2 core recombination site"
     primer_bind     3017..3065
                     /standard_name="P3_F2"
     primer_bind     complement(3017..3065)
                     /standard_name="P3_R2"
     primer_bind     3017..3031
                     /standard_name="15 bp EcoRV Overhang"
     misc_feature    3032..3037
                     /standard_name="BsaI"
     misc_feature    3032..3050
                     /note="Geneious type: Editing History Insertion"
                     /standard_name
     misc_feature    complement(3044..3050)
                     /standard_name="SapI"
     primer_bind     3051..3065
                     /standard_name="15 bp EcoRV Overhang"
     promoter        complement(3055..3073)
                     /note="Geneious type: Promoter"
                     /standard_name="T7 prom"
     primer_bind     complement(3078..3094)
                     /standard_name="M13 Reverse (Quintara)"
     CDS             3207..4016
                     /standard_name="Kanamycin resistance gene"
     rep_origin      4137..4810
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 atagaaggca cagtcgaggg actggagacc
      601 ccaaataatg attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgatgagca
      661 atgctttttt ataatgccaa ctttgtacaa aaaagctgaa cgagaaacgt aaaatgatat
      721 aaatatcaat atattaaatt agattttgca taaaaaacag actacataat actgtaaaac
      781 acaacatatc cagtcactat gaatcaacta cttagatggt attagtgacc tgtagtcgac
      841 cgacagcctt ccaaatgttc ttcgggtgat gctgccaact tagtcgaccg acagccttcc
      901 aaatgttctt ctcaaacgga atcgtcgtat ccagcctact cgctattgtc ctcaatgccg
      961 tattaaatca taaaaagaaa taagaaaaag aggtgcgagc ctcttttttg tgtgacaaaa
     1021 taaaaacatc tacctattca tatacgctag tgtcatagtc ctgaaaatca tctgcatcaa
     1081 gaacaatttc acaactctta tacttttctc ttacaagtcg ttcggcttca tctggatttt
     1141 cagcctctat acttactaaa cgtgataaag tttctgtaat ttctactgta tcgacctgca
     1201 gactggctgt gtataaggga gcctgacatt tatattcccc agaacatcag gttaatggcg
     1261 tttttgatgt cattttcgcg gtggctgaga tcagccactt cttccccgat aacggagacc
     1321 ggcacactgg ccatatcggt ggtcatcatg cgccagcttt catccccgat atgcaccacc
     1381 gggtaaagtt cacgggagac tttatctgac agcagacgtg cactggccag ggggatcacc
     1441 atccgtcgcc cgggcgtgtc aataatatca ctctgtacat ccacaaacag acgataacgg
     1501 ctctctcttt tataggtgta aaccttaaac tgcatttcac cagcccctgt tctcgtcagc
     1561 aaaagagccg ttcatttcaa taaaccgggc gacctcagcc atcccttcct gattttccgc
     1621 tttccagcgt tcggcacgca gacgacgggc ttcattctgc atggttgtgc ttaccagacc
     1681 ggagatattg acatcatata tgccttgagc aactgatagc tgtcgctgtc aactgtcact
     1741 gtaatacgct gcttcatagc atacctcttt ttgacatact tcgggtatac atatcagtat
     1801 atattcttat accgcaaaaa tcagcgcgca aatacgcata ctgttatctg gcttttagta
     1861 agccggatcc acgcggcgtt tacgcccccc ctgccactca tcgcagtact gttgtaattc
     1921 attaagcatt ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag
     1981 cggcatcagc accttgtcgc cttgcgtata atatttgccc atggtgaaaa cgggggcgaa
     2041 gaagttgtcc atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc
     2101 tgagacgaaa aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta
     2161 acacgccaca tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact
     2221 ccagagcgat gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact
     2281 atcccatatc accagctcac cgtctttcat tgccatacgg aattccggat gagcattcat
     2341 caggcgggca agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt
     2401 ctttaaaaag gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga
     2461 ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc
     2521 agtgattttt ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa
     2581 tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgccgatc
     2641 aacgtctcat tttcgccaaa agttggccca gggcttcccg gtatcaacag ggacaccagg
     2701 atttatttat tctgcgaagt gatcttccgt cacaggtatt tattcggcgc aaagtgcgtc
     2761 gggtgatgct gccaacttag tcgactacag gtcactaata ccatctaagt agttgattca
     2821 tagtgactgg atatgttgtg ttttacagta ttatgtagtc tgttttttat gcaaaatcta
     2881 atttaatata ttgatattta tatcatttta cgtttctcgt tcagctttct tgtacaaagt
     2941 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     3001 aaataaaatc attatttgcc atccagctga tggtctcctt cacgaagagc atcccctata
     3061 gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc
     3121 tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt
     3181 acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc
     3241 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
     3301 ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc
     3361 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
     3421 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
     3481 catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc
     3541 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga
     3601 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat
     3661 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
     3721 ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg
     3781 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
     3841 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
     3901 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
     3961 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag
     4021 aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa
     4081 gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg
     4141 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4201 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4261 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4321 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4381 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4441 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4501 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4561 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4621 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4681 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4741 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4801 ttgctcacat gtt
//
